Secreted protein HEMCM42

ABSTRACT

The present invention relates to novel human secreted proteins and isolated nucleic acids containing the coding regions of the genes encoding such proteins. Also provided are vectors, host cells, antibodies, and recombinant methods for producing human secreted proteins. The invention further relates to diagnostic and therapeutic methods useful for diagnosing and treating disorders related to these novel human secreted proteins.

FIELD OF THE INVENTION

[0001] This application is a continuation-in-part of, and claims benefitunder 35 U.S.C. § 120 of copending U.S. patent application Ser. No:PCT/US98/10868, filed May 28, 1998, which is hereby incorporated byreference, which claims benefit under 35 U.S.C. § 119(e) based on U.S.Provisional Applications: Filing Date Appln No. 1. May 30, 1997 60/044,039 2. May 30, 1997 60/048, 093 3. May 30, 1997 60/048, 190 4. May 30,1997 60/050, 935 5. May 30, 1997 60/048, 101 6. May 30, 1997 60/048, 3567. Aug. 29, 1997 60/056, 250 8. Aug. 29, 1997 60/056, 296 9. Aug. 29,1997 60/056, 293

[0002] which are hereby incorporated by reference.

[0003] This invention relates to newly identified polynucleotides andthe polypeptides encoded by these polynucleotides, uses of suchpolynucleotides and polypeptides, and their production.

BACKGROUND OF THE INVENTION

[0004] Unlike bacterium, which exist as a single compartment surroundedby a membrane, human cells and other eucaryotes are subdivided bymembranes into many functionally distinct compartments. Eachmembrane-bounded compartment, or organelle, contains different proteinsessential for the function of the organelle. The cell uses “sortingsignals,” which are amino acid motifs located within the protein, totarget proteins to particular cellular organelles.

[0005] One type of sorting signal, called a signal sequence, a signalpeptide, or a leader sequence, directs a class of proteins to anorganelle called the endoplasmic reticulum (ER). The ER separates themembrane-bounded proteins from all other types of proteins. Oncelocalized to the ER, both groups of proteins can be further directed toanother organelle called the Golgi apparatus. Here, the Golgidistributes the proteins to vesicles, including secretory vesicles, thecell membrane, lysosomes, and the other organelles.

[0006] Proteins targeted to the ER by a signal sequence can be releasedinto the extracellular space as a secreted protein. For example,vesicles containing secreted proteins can fuse with the cell membraneand release their contents into the extracellular space—a process calledexocytosis. Exocytosis can occur constitutively or after receipt of atriggering signal. In the latter case, the proteins are stored insecretory vesicles (or secretory granules) until exocytosis istriggered. Similarly, proteins residing on the cell membrane can also besecreted into the extracellular space by proteolytic cleavage of a“linker” holding the protein to the membrane.

[0007] Despite the great progress made in recent years, only a smallnumber of genes encoding human secreted proteins have been identified.These secreted proteins include the commercially valuable human insulin,interferon, Factor VIII, human growth hormone, tissue plasminogenactivator, and erythropoeitin. Thus, in light of the pervasive role ofsecreted proteins in human physiology, a need exists for identifying andcharacterizing novel human secreted proteins and the genes that encodethem. This knowledge will allow one to detect, to treat, and to preventmedical disorders by using secreted proteins or the genes that encodethem.

SUMMARY OF THE INVENTION

[0008] The present invention relates to novel polynucleotides and theencoded polypeptides. Moreover, the present invention relates tovectors, host cells, antibodies, and recombinant methods for producingthe polypeptides and polynucleotides. Also provided are diagnosticmethods for detecting disorders related to the polypeptides, andtherapeutic methods for treating such disorders. The invention furtherrelates to screening methods for identifying binding partners of thepolypeptides.

DETAILED DESCRIPTION

[0009] Definitions

[0010] The following definitions are provided to facilitateunderstanding of certain terms used throughout this specification.

[0011] In the present invention, “isolated” refers to material removedfrom its original environment (e.g., the natural environment if it isnaturally occurring), and thus is altered “by the hand of man” from itsnatural state. For example, an isolated polynucleotide could be part ofa vector or a composition of matter, or could be contained within acell, and still be “isolated” because that vector, composition ofmatter, or particular cell is not the original environment of thepolynucleotide.

[0012] In the present invention, a “secreted” protein refers to thoseproteins capable of being directed to the ER, secretory vesicles, or theextracellular space as a result of a signal sequence, as well as thoseproteins released into the extracellular space without necessarilycontaining a signal sequence. If the secreted protein is released intothe extracellular space, the secreted protein can undergo extracellularprocessing to produce a “mature” protein. Release into the extracellularspace can occur by many mechanisms, including exocytosis and proteolyticcleavage.

[0013] As used herein, a “polynucleotide” refers to a molecule having anucleic acid sequence contained in SEQ ID NO:X or the cDNA containedwithin the clone deposited with the ATCC. For example, thepolynucleotide can contain the nucleotide sequence of the full lengthcDNA sequence, including the 5′ and 3′ untranslated sequences, thecoding region, with or without the signal sequence, the secreted proteincoding region, as well as fragments, epitopes, domains, and variants ofthe nucleic acid sequence. Moreover, as used herein, a “polypeptide”refers to a molecule having the translated amino acid sequence generatedfrom the polynucleotide as broadly defined.

[0014] In the present invention, the full length sequence identified asSEQ ID NO:X was often generated by overlapping sequences contained inmultiple clones. (contig analysis). A representative clone containingall or most of the sequence for SEQ ID NO:X was deposited with theAmerican Type Culture Collection (“ATCC”). As shown in Table 1, eachclone is identified by a cDNA Clone ID (Identifier) and the ATCC DepositNumber. The ATCC is located at 10801 University Boulevard, Manassas, Va.20110-2209, USA. The ATCC deposit was made pursuant to the terms of theBudapest Treaty on the international recognition of the deposit ofmicroorganisms for purposes of patent procedure.

[0015] A “polynucleotide” of the present invention also includes thosepolynucleotides capable of hybridizing, under stringent hybridizationconditions, to sequences contained in SEQ ID NO:X, the complementthereof, or the cDNA within the clone deposited with the ATCC.“Stringent hybridization conditions” refers to an overnight incubationat 42° C. in a solution comprising 50% formamide, 5× SSC (750 mM NaCl,75 mM sodium citrate), 50 mM sodium phosphate (pH 7.6), 5× Denhardt'ssolution, 10% dextran sulfate, and 20 μg/ml denatured, sheared salmonsperm DNA, followed by washing the filters in 0.1×SSC at about 65° C.

[0016] Also contemplated are nucleic acid molecules that hybridize tothe polynucleotides of the present invention at lower stringencyhybridization conditions. Changes in the stringency of hybridization andsignal detection are primarily accomplished through the manipulation offormamide concentration (lower percentages of formanide result inlowered stringency); salt conditions, or temperature. For example, lowerstringency conditions include an overnight incubation at 37° C. in asolution comprising 6× SSPE (20× SSPE=3M NaCl; 0.2M NaH₂PO₄; 0.02M EDTA,pH 7.4), 0.5% SDS, 30% formamide, 100 ug/ml salmon sperm blocking DNA;followed by washes at 50° C. with 1×SSPE, 0.1% SDS. In addition, toachieve even lower stringency, washes performed following stringenthybridization can be done at higher salt concentrations (e.g. 5× SSC).

[0017] Note that variations in the above conditions may be accomplishedthrough the inclusion and/or substitution of alternate blocking reagentsused to suppress background in hybridization experiments. Typicalblocking reagents include Denhardt's reagent, BLOTTO, heparin, denaturedsalmon sperm DNA, and commercially available proprietary formulations.The inclusion of specific blocking reagents may require modification ofthe hybridization conditions described above, due to problems withcompatibility.

[0018] Of course, a polynucleotide which hybridizes only to polyA+sequences (such as any 3′ terminal polyA+ tract of a cDNA shown in thesequence listing), or to a complementary stretch of T (or U) residues,would not be included in the definition of “polynucleotide,” since sucha polynucleotide would hybridize to any nucleic acid molecule containinga poly (A) stretch or the complement thereof (e.g., practically anydouble-stranded cDNA clone).

[0019] The polynucleotide of the present invention can be composed ofany polyribonucleotide or polydeoxribonucleotide, which may beunmodified RNA or DNA or modified RNA or DNA. For example,polynucleotides can be composed of single- and double-stranded DNA, DNAthat is a mixture of single- and double-stranded regions, single- anddouble-stranded RNA, and RNA that is mixture of single- anddouble-stranded regions, hybrid molecules comprising DNA and RNA thatmay be single-stranded or, more typically, double-stranded or a mixtureof single- and double-stranded regions. In addition, the polynucleotidecan be composed of triple-stranded regions comprising RNA or DNA or bothRNA and DNA. A polynucleotide may also contain one or more modifiedbases or DNA or RNA backbones modified for stability or for otherreasons. “Modified” bases include, for example, tritylated bases andunusual bases such as inosine. A variety of modifications can be made toDNA and RNA; thus, “polynucleotide” embraces chemically, enzymatically,or metabolically modified forms.

[0020] The polypeptide of the present invention can be composed of aminoacids joined to each other by peptide bonds or modified peptide bonds,i.e., peptide isosteres, and may contain amino acids other than the 20gene-encoded amino acids. The polypeptides may be modified by eithernatural processes, such as posttranslational processing, or by chemicalmodification techniques which are well known in the art. Suchmodifications are well described in basic texts and in more detailedmonographs, as well as in a voluminous research literature.Modifications can occur anywhere in a polypeptide, including the peptidebackbone, the amino acid side-chains and the amino or carboxyl termini.It will be appreciated that the same type of modification may be presentin the same or varying degrees at several sites in a given polypeptide.Also, a given polypeptide may contain many types of modifications.Polypeptides may be branched , for example, as a result ofubiquitination, and they may be cyclic, with or without branching.Cyclic, branched, and branched cyclic polypeptides may result fromposttranslation natural processes or may be made by synthetic methods.Modifications include acetylation, acylation, ADP-ribosylation,amidation, covalent attachment of flavin, covalent attachment of a hememoiety, covalent attachment of a nucleotide or nucleotide derivative,covalent attachment of a lipid or lipid derivative, covalent attachmentof phosphotidylinositol, cross-linking, cyclization, disulfide bondformation, demethylation, formation of covalent cross-links, formationof cysteine, formation of pyroglutamate, formylation,gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation,iodination, methylation, myristoylation, oxidation, pegylation,proteolytic processing, phosphorylation, prenylation, racemization,selenoylation, sulfation, transfer-RNA mediated addition of amino acidsto proteins such as arginylation, and ubiquitination. (See, forinstance, PROTEINS—STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E.Creighton, W. H. Freeman and Company, New York (1993); POSTTRANSLATIONALCOVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press,New York, pgs. 1-12 (1983); Seifter et al., Meth Enzymol 182:626-646(1990); Rattan et al., Ann NY Acad Sci 663:48-62 (1992).)

[0021] “SEQ ID NO:X” refers to a polynucleotide sequence while “SEQ IDNO:Y” refers to a polypeptide sequence, both sequences identified by aninteger specified in Table 1.

[0022] “A polypeptide having biological activity” refers to polypeptidesexhibiting activity similar, but not necessarily identical to, anactivity of a polypeptide of the present invention, including matureforms, as measured in a particular biological assay, with or withoutdose dependency. In the case where dose dependency does exist, it neednot be identical to that of the polypeptide, but rather substantiallysimilar to the dose-dependence in a given activity as compared to thepolypeptide of the present invention (i.e., the candidate polypeptidewill exhibit greater activity or not more than about 25-fold less and,preferably, not more than about tenfold less activity, and mostpreferably, not more than about three-fold less activity relative to thepolypeptide of the present invention.)

[0023] Polynucleotides and Polypeptides of the Invention

[0024] FEATURES OF PROTEIN ENCODED BY GENE NO: 1

[0025] This gene maps to chromosome 3 and therefore polynucleotides ofthe present invention can be used in linkage analysis as a marker forchromosome 3.

[0026] This gene is expressed in several fetal tissues including brain,liver, and lung, and to a lesser extent, in adult tissues, particularlyskin.

[0027] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,developmental, neural, hepatic, or pulmonary disorders, particularycancers of the above system and tissues. Similarly, polypeptides andantibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the central nervous system, hepatic system and hepaticsystem, expression of this gene at significantly higher or lower levelsmay be routinely detected in certain tissues or cell types (e.g. brain,cancerous and wounded tissues) or bodily fluids (e.g., lymph, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0028] The tissue distribution indicates that polynucleotides andpolypeptides corresponding to this gene are useful for serving as atarget for a variety of blocking agents, as it is likely to be involvedin the promotion of a variety of cancers. Furthermore, polynucleotidesand polypeptides corresponding to this gene are useful for the diagnosisand treatment of cancer and other proliferative disorders. Expressionwithin embryonic tissue and other cellular sources marked byproliferating cells indicates that this protein may play a role in theregulation of cellular division. Additionally, the expression inhematopoietic cells and tissues indicates that this protein may play arole in the proliferation, differentiation, and/or survival ofhematopoietic cell lineages. In such an event, this gene may be usefulin the treatment of lymphoproliferative disorders, and in the-maintenance and differentiation of various hematopoietic lineages fromearly hematopoietic stem and committed progenitor cells. Similarly,embryonic development also involves decisions involving celldifferentiation and/or apoptosis in pattern formation. Thus this proteinmay also be involved in apoptosis or tissue differentiation and couldagain be useful in cancer therapy. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0029] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:11 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1155 of SEQID NO:11, b is an integer of 15 to 1169, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:11, and whereb is greater than or equal to a+14.

[0030] FEATURES-OF PROTEIN ENCODED BY GENE NO: 2

[0031] In specific embodiments, the polypeptides of the inventioncomprise the sequence:MSVPAFIDISEEDQAAELRAYLKSKGAEISEENSEGGLHVDLAQIIEACDVCLKEDDKDVESVMNSVVSLLLILEPDKQEALIESLCEKLVKFREGERPSLRLQLLSNLFHGMDKNTPVRYTVYCSLIKVAASCGAIQYIPTELDQVRKWISDWNLTTEKKHTLLRLLYEALVDCKKSDAASKVMVELLGSYTEDNASQARVDAHRCIVRALKDPNAFLFDHLLTLKPVKFLEGELIHDLLTIFVSAKLASYVKFYQNNKDFIDSLGLLHEQNMAKMRLLTFMGMAVENKEISFDTMQQELQIGADDVEAFVIDAVRTKMVYCKIDQTQRKVVVSHSTHRTFGKQQWQQLYDTLNAWKQNLNKVKNSLLS LSDT (SEQ ID NO:83),MSVPAFIDISEED (SEQ ID NO:84), QAAELRAYLKS KGAE (SEQ ID NO:85),ISEENSEGGLHVDLAQI (SEQ ID NO:86), IEACDVCLK EDDKDVESV (SEQ ID NO:87),VARPSSLFRSAWSCEW (SEQ ID NO:88), LRL QLLSNLFHG (SEQ ID NO:89),KDVESVMNSVVSLLLIL (SEQ ID NO:90), DAAS KVMVELLGSYTEDNASQARVDA (SEQ IDNO:91), KMRLLTFMGMAVENKEIS (SEQ ID NO:93), and/or VEAFVIDAVR (SEQ IDNO:92). Polynucleotides encoding these polypeptides are also encompassedby the invention. The translation product of this gene shares homologywith the Homo sapiens GA17 protein (gb accession number AF064603). Thegene encoding the disclosed cDNA is thought to reside on the Xchromosome. Accordingly, polynucleotides related to this invention areuseful as a marker in linkage analysis for the X chromosome.

[0032] This gene is expressed primarily in bone, and to a lesser extent,in brain, lung, T-cells, muscle, skin, testis, spleen and macrophages.

[0033] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, skeletal,neural, or hematopoietic disorders, particularly bone cancer,osteoarthritis, autoimmune diseases. Similarly, polypeptides andantibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the immune system and skeletal system, expression ofthis gene at significantly higher or lower levels may be routinelydetected in certain tissues or cell types (e.g. bone, brain, lung,testis, and cancerous and wounded tissues) or bodily fluids (e.g.,lymph, pulmonary surfactant or sputum, serum, plasma, urine, synoviafluid or spinal fluid) or another tissue or cell sample taken from anindividual having such a disorder, relative to the standard geneexpression level, i.e., the expression level in healthy tissue or bodilyfluid from an individual not having the disorder.

[0034] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:48 as residues: Arg-31 to Ser-37, Met-50 to Val-56, Glu-80 toTrp-87, Thr-94 to His-99, Tyr-129 to Ser-135, Tyr-193 to Phe-199,Ser-274 to Gln-285, Ala-293 to Lys-302.

[0035] The tissue distribution in bone and immune cells and tissuesindicates that polynucleotides and polypeptides corresponding to thisgene are useful for the diagnosis and/or treatment of skeletal andhematopoietic disorders. Elevated levels of expression of this geneproduct in bone suggest that it may be useful in influencing bone massin such conditions as osteoporosis, or in the stimulation or growth ofbone, cartilage, and/or tendons, for example. More generally, asevidenced by expression in T-cells and spleen, this gene may play a rolein the survival, proliferation, and/or differentiation of hematopoieticcells in general, and may be of use in augmentation of the numbers ofstem cells and committed progenitors. Expression of this gene product inmacrophage also indicates that it may play a role in mediating responsesto infection and controlling immunological responses, such as those thatoccur during immune surveillance. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0036] Many polynticleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:12 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1296 of SEQID NO:12, b is an integer of 15 to 1310, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:12, and whereb is greater than or equal to a+14.

[0037] FEATURES OF PROTEIN ENCODED BY GENE NO: 3

[0038] The translation product of this gene shares sequence homologywith various kinases. The closest homolog is the mouse TIF1 which is aconserved nuclear protein. TIF1 enhances RXR and RAR AF-2 in yeast andinteracts in a ligand-dependent manner with several nuclear receptors inyeast and mammalian cells, as well as in vitro. Remarkably, theseinteractions require the amino acids constituting the AF-2 activatingdomain conserved in all active NRs. Moreover, the estrogen receptor (ER)AF-2 antagonist hydroxytamoxifen cannot promote ER-TIF1 interaction.TIF1, which contains several conserved domains found in transcriptionalregulatory proteins, may be a mediator of ligand-dependent AF-2.Interestingly, the TIF1 N-terminal moiety is fused to B-raf in the mouseoncoprotein T18. One embodiment of this gene comprises polypeptides ofthe following amino acid sequence:MEAVPEGDWFCTVCLAQQVEGEFTQKPGFPKRGQKRKSGYSLNFSEGDGRRR RVLLRGRESPAAGPRYSEEGLSPSKRRRLSMRNHHSDLTFCEIILMEMESHD AAWPFLEPVNPRLVSGYRRIIKNPMDFSTMRERLLRGGYTSSEEFAADALLV FDNCQTFNEDDSEVGKAGHIMRRFFESRWEEFYQGKQANL(SEQ ID NO:94), MEAVPEGDWFCTVCLAQQVE (SEQ ID NO:95), GEFTQKPGFPKRGQKRKSGYS (SEQ ID NO:96), LNFSEGDGRRRRVLLRGRESP (SEQ ID NO:97), AAGPRYSEEGLSPSKRRRLS (SEQ ID NO:98), MRNHHSDLTFCEIILMEMESH (SEQ ID NO:99),DAAWPFLEPVNPRLVSGYRR (SEQ ID NO:100), IIKNPMDFSTMR ERLLRGGYT (SEQ IDNO:101), SSEEFAADALLVFDNCQTFNE (SEQ ID NO: 102), DDSEVGKAGHIMRRFFE (SEQID NO:103), and/or SRWEEFYQGKQA NL (SEQ ID NO:104). An additionalembodiment is the polynucleotides encoding these polypeptides.

[0039] This gene is expressed primarily in activated T-cells, and to alesser extent, in testes and brain.

[0040] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune,reproductive, or neural disorders, particularly autoimmune diseases,AIDS, leukemias, and cancers. Similarly, polypeptides and antibodiesdirected to these polypeptides are useful in providing immunologicalprobes for differential identification of the tissue(s) or cell type(s).For a number of disorders of the above tissues or cells, particularly ofthe immune, reproductive, or neural systems, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. immune, neural, reproductive, andcancerous and wounded tissues) or bodily fluids (e.g., lymph, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in Healthytissue or bodily fluid from an individual not having the disorder.

[0041] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:49 as residues: Ala-31 to Glu-36.

[0042] The tissue distribution in activated T-cells, combined with thehomology to TIF indicates that polynucleotides and polypeptidescorresponding to this gene are useful for modulation of nuclear receptorand ligand interaction in various immune disorders. Furthermore, thesecreted protein can also be used to determine biological activity, toraise antibodies, as tissue markers, to isolate cognate ligands orreceptors, to identify agents that modulate their interactions and asnutritional supplements. It may also have a very wide range ofbiological acitivities. Typical of these are cytokine, cellproliferation/differentiation modulating activity or induction of othercytokines; immunostimulating/immunosuppressant activities (e.g. fortreating human immunodeficiency virus infection, cancer, autoimmunediseases and allergy); regulation of hematopoiesis (e.g. for treatinganaemia or as adjunct to chemotherapy); stimulation or growth of bone,cartilage, tendons, ligaments and/or nerves (e.g. for treating wounds,stimulation of follicle stimulating hormone (for control of fertility);chemotactic and chemokinetic activities (e.g. for treating infections,tumors); hemostatic or thrombolytic activity (e.g. for treatinghaemophilia, cardiac infarction etc.); anti-inflammatory activity (e.g.for treating septic shock, Crohn's disease); as antimicrobials; fortreating psoriasis or other hyperproliferative diseases; for regulationof metabolism, and behaviour. Also contemplated is the use of thecorresponding nucleic acid in gene therapy procedures. Protein, as wellas, antibodies directed against the protein may show utility as a tumormarker and/or immunotherapy targets for the above listed tissues.

[0043] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:13 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1125 of SEQID NO:13, b is an integer of 15 to 1139, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:13, and whereb is greater than or equal to a+14.

[0044] FEATURES OF PROTEIN ENCODED BY GENE NO: 4

[0045] This gene maps to chromosome 11. Accordingly, polynucleotides ofthe invention can be used in linkage analysis as a marker for chromosome11. In specific embodiments, the polypeptides of the invention comprisethe sequence: MSEIYLRCQDEQQYARWMAGCRLASKGRTMADSSY (SEQ ID NO:105), LVAPRFQRKFKAKQLTPRILEAHQNVAQLSLAEAQLFIQAWQSL (SEQ ID NO:106),VGDVVKTWRFSNMRQWNVNWDIR (SEQ ID NO:107), EEIDCTEEEMMVFA ALQYHINKLSQS(SEQ ID NO:108), and/or EEIDCTEEEMMVFAALQYHINKLS QS (SEQ ID NO:109).Polynucleotides encoding these polypeptides are also encompassed by theinvention.

[0046] This gene is expressed primarily in white blood cell typesincluding monocytes, T-cells, neutrophils, and to a lesser extent, inumbilical vein and liver.

[0047] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, variousdiseases of the immune system including AIDS, immunodeficency diseases,and autoimmune disorders, in addition to hepatic and developmentaldisorders. Similarly, polypeptides and antibodies directed to thesepolypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theimmune or hepatic system, expression of this gene at significantlyhigher or lower levels may be routinely detected in certain tissues orcell types (e.g. immune, hepatic, developmental, and cancerous andwounded tissues) or bodily fluids (e.g., lymph, bile, serum, plasma,urine, synovial fluid or spinal fluid) or another tissue or cell sampletaken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0048] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:50 as residues: Ser-3 to Pro-9, Leu-17 to Leu-29, Asp-64 toPro-69, Ile-105 to Gln-110, Thr-183 to Gln-200, Cys-239 to Arg-247,Ser-256 to Met-261, Gln-280 to Ala-296, Arg-310 to Thr-321, Lys-363 toAsp-368, Ser-395 to Trp-400, Thr-443 to Asp-453.

[0049] The tissue distribution in a variety of immune cells and tissuesindicates that polynucleotides and polypeptides corresponding to thisgene are useful for replacement therapy in a variety of immune systemdisorders. Furthermore, polynucleotides and polypeptides correspondingto this gene are useful for the diagnosis and/or treatment ofhematopoietic disorders. This gene product is primarily expressed inhematopoietic cells and tissues, suggesting that it plays a role in thesurvival, proliferation, and/or differentiation of hematopoieiticlineages. This is particularly supported by the expression of this geneproduct in liver, which is a primary site of definitive hematopoiesisduring development. Expression of this gene product in T cells andprimary dendritic cells also strongly indicates a role for this proteinin immune function and immune surveillance. Protein, as well as,antibodies directed against the protein may show utility as a tumormarker and/or immunotherapy targets for the above listed tissues.

[0050] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:14 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 2257 of SEQID NO:14, b is an integer of 15 to 2271, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:14, and whereb is greater than or equal to a+14.

[0051] FEATURES OF PROTEIN ENCODED BY GENE NO: 5

[0052] The gene encoding the disclosed cDNA is thought to reside onchromosome 15. Accordingly, polynucleotides related to this inventionare useful as a marker in linkage analysis for chromosome 15.

[0053] This gene is expressed primarily in brain.

[0054] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, neuraldisorders, particularly neurodegenerative and behaviour conditions, suchas mood disorders, schizophrenia and related diseases, bipolar disorderand unipolar depression. Similarly, polypeptides and antibodies directedto these polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of thecentral nervous system, expression of this gene at significantly higheror lower levels may be routinely detected in certain tissues or celltypes (e.g. brain, and cancerous and wounded tissues) or bodily fluids(e.g., lymph, serum, plasma, urine, synovial fluid or spinal fluid) oranother tissue or cell sample taken from an individual having such adisorder, relative to the standard gene expression level, i.e., theexpression level in healthy tissue or bodily fluid from an individualnot having the disorder.

[0055] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:51 as residues: Met-1 to Gly-8, Pro-10 to Arg-17, Pro-45 toSer-55, Gly-63 to Tyr-74.

[0056] The tissue distribution in brain indicates that polynucleotidesand polypeptides corresponding to this gene are useful for thedetection/treatment of neurodegenerative disease states and behaviouraldisorders such as Alzheimers Disease, Parkinsons Disease, HuntingtonsDisease, schizophrenia, mania, dementia, paranoia, obsessive compulsivedisorder and panic disorder. Also given the brain-specific expression ofthis gene, the promoter region of this gene may contain a brain-specificelement that could be used for targeting expression of vector systems tothe brain in gene replacement therapy. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues

[0057] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:15 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral fornula of a-b, where a is any integer between 1 to 612 of SEQID NO:15, b is an integer of 15 to 626, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:15, and where bis greater than or equal to a+14.

[0058] FEATURES OF PROTEIN ENCODED BY GENE NO: 6

[0059] This gene maps to chromosome 1 and therefore, polynucleotides ofthe invention can be used in linkage analysis as a marker for chromosome1.

[0060] This gene is expressed abundantly in rhabdomyosarcoma, and isexpressed to a high level indifferent regions of the brain and pituitarygland.

[0061] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,neurological, muscular, or endocrine disorders, particularly soft tissuetumors, such as fibroids. Similarly, polypeptides and antibodiesdirected to these polypeptides are useful in providing immunologicalprobes for differential identification of the tissue(s) or cell type(s).For a number of disorders of the above tissues or cells, particularly ofthe brain, expression of this gene at significantly higher or lowerlevels may be routinely detected in certain tissues or cell types (e.g.brain, muscle, endocrine, and cancerous and wounded tissues) or bodilyfluids (e.g., lymph, serum, plasma, urine, synovial fluid or spinalfluid) or another tissue or cell sample taken from an individual havingsuch a disorder, relative to the standard gene expression level, i.e.,the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0062] The tissue distribution in rhadomyosarcoma tissue indicates arole for the protein product either in the detection and/or treatment ofmusculo-skeletal disorders including muscle degeneration, musclewasting, rhabdomyolysis, muscular dystrophy, cardiomyopathy, fibroids,myomas, and rhabdomyosarcomas. Furthermore, expression in the brainindicates a role for the protein product of this gene in thedetection/treatment of neurodegenerative disease states and behaviouraldisorders such as Alzheimers Disease, Parkinsons Disease, HuntingtonsDisease, schizophrenia, mania, dementia, paranoia, obsessive compulsivedisorder and panic disorder. Protein, as well as, antibodies directedagainst the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0063] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:16 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 2104 of SEQID NO:16, is an integer of 15 to 2118, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:16, and where bis greater than or equal to a+14.

[0064] FEATURES OF PROTEIN ENCODED BY GENE NO: 7

[0065] The translation product of this gene shares sequence homologywith the TDAG51 gene which is thought to be important in the mediationof apoptosis and cell death by coupling TCR stimulation to Fasexpression. In specific embodiments, the polypeptides of the inventioncomprise the sequence: KELSFARIKAVECVESTGRHIYFTLV (SEQ ID NO:110) and/orGWNAQITLGLV KFKNQQ (SEQ ID NO:111). The gene encoding the disclosed cDNAis thought to reside on chromosome 1. Accordingly, polynucleotidesrelated to this invention are useful as a marker in linkage analysis forchromosome 1.

[0066] This gene is expressed in breast cancer tissue, and to a lesserextent, in macrophage.

[0067] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune orreproductive disorders, particularly breast cancer. Similarly,polypeptides and antibodies directed to these polypeptides are useful inproviding immunological probes for differential identification of thetissue(s) or cell type(s). For a number of disorders of the abovetissues or cells, particularly of the immune system, expression of thisgene at significantly higher or lower levels may be routinely detectedin certain tissues or cell types (e.g. breast, immune, and cancerous andwounded tissues) or bodily fluids (e.g., lymph, breast milk, serum,plasma, urine, synovial fluid inal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0068] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:54 as residues: Met-1 to Pro-9, Gln-43 to Glu-49, Phe-95 toArg-102.

[0069] The tissue distribution in breast cancer tissue, combined withthe homology to the TDAG51 gene indicates that polynucleotides andpolypeptides corresponding to this gene are useful for diagnosis andintervention of immune disorders, such as immunodeficiency, allergy,infection, inflammation, and tissue/organ transplantation.Alternatively, the tissue distribution in tumors of breast originsindicates that polynucleotides and polypeptides corresponding to thisgene are useful for the diagnosis and intervention of these tumors,particularly during antigen presentation early in the immune response.Protein, as well as, antibodies directed against the protein may showutility as a tissue-specific marker and/or immunotherapy target for theabove listed tissues.

[0070] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:17 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1062 of SEQID NO:17, b is an integer of 15 to 1076, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:17, and whereb is greater than or equal to a+14.

[0071] FEATURES OF PROTEIN ENCODED BY GENE NO: 8

[0072] The gene encoding the disclosed cDNA is thought to reside onchromosome 1. Accordingly, polynucleotides related to this invention areuseful as a marker in linkage analysis for chromosome 1. One embodimentof this gene comprises the polypeptides of the following amino acidsequence: (SEQ ID NO:112)MVTTIVLGRRFIGSIVKEASQRGKVSLFRSILLFLTRFTVLTATGWSLCRSLIHLFRTYSFLNLLFLCYPFGMYIPFLQLNXXLRKTSLFNHMASMGPRE AVSGLAKSRDYLLTLRETWKQHXRQLYGPDAMPTHACCLSPSLIRSEVE FLKMDFNWRMKEVLVSSMLSAYYVAFYPVWFVKNTHYYDKRWSCXTLPA GVHQHLRDPHAAPAACQLL, (SEQ ID NO:113) MVTTIVLGRRFIGSIVKEASQRGKVS,(SEQ ID NO:114) LFRSILLFLTRFTVLTATGWSLC, (SEQ ID NO:115) RSLIHLFRTYSFLNLLFLCYPFGMYIPFLQ, (SEQ ID NO:116) LNXXLRKTSLFNHMASMGPREAVSGLAKSR, (SEQ ID NO:117) DYLLTLRETWKQHXRQLYGPD AMPTHACCL, (SEQID NO:118) SPSLIRSEVEFLKMDFNWRMKEVLVSSMLSA, (SEQ ID NO:119)YYVAFVPVWFVKNTHYYDKRWSCXTLP, and/or (SEQ ID NO:120)AGVHQHLRDPHAAPAACQLL.

[0073] An additional embodiment is the polynucleotides encoding thesepolypeptides.

[0074] This gene is expressed in breast tissue, amiotic cells, and to alesser extent, in smooth muscle, T-cells, and infant brain.

[0075] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,reproductive or developmental disorders, particularly fetal distresssyndrome and embryonic wasting. Similarly, polypeptides and antibodiesdirected to these polypeptides are useful in providing immunologicalprobes for differential identification of the tissue(s) or cell type(s).For a number of disorders of the above tissues or cells, particularly ofthe female reproductive system, expression of this gene at significantlyhigher or lower levels may be routinely detected in certain tissues orcell types (e.g. breast, fetal, neural, immune, hematopoietic, muscular,and cancerous and wounded tissues) or bodily fluids (e.g., lymph,amniotic fluid, serum, plasma, urine, synovial fluid or spinal fluid) oranother tissue or cell sample taken from an individual having such adisorder, relative to the standard gene expression level, i.e., theexpression level in healthy tissue or bodily fluid from an individualnot having the disorder.

[0076] The tissue distribution in breast tissue indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the diagnosis and/or treatment of reproductive disorders.Alternately, this gene product may be produced by the placenta and thentransported to the embryo, where it may play a crucial role in thedevelopment and/or survival of the developing embryo or fetus.Expression of this gene product in a vascular-rich tissue such as thebreast and amnion also indicates that this gene product may be producedmore generally in endothelial cells or within the circulation. In suchinstances, it may play more generalized roles in vascular function, suchas in angiogenesis. It may also be produced in the vasculature and haveeffects on other cells within the circulation, such as hematopoieticcells. It may serve to promote the proliferation, survival, activation,and/or differentiation of hematopoietic cells, as well as other cellsthroughout the body. Protein, as well as, antibodies directed againstthe protein may show utility as a tumor marker and/or immunotherapytargets for the above listed tissues.

[0077] FEATURES OF PROTEIN ENCODED BY GENE NO: 9

[0078] In specific embodiments, the polypeptides of the inventioncomprise the sequence: LVLGLSXLNNSYNFSF (SEQ ID NO:121),HVVIGSQAEEGQYSLNF (SEQ ID NO:122), HNCNNSVPGKEHPFDITVM (SEQ ID NO:123),FIKYVLSD KEKKVFGIV (SEQ ID NO:124), IPMQVLANVAYII (SEQ ID NO:125),IPMQVL ANVAYII (SEQ ID NO:126), DGKVAVNLAKLKLFR (SEQ ID NO:127), and/orIREKNPDGFLSAA (SEQ ID NO:128). Polynucleotides encoding thesepolypepides are also encompassed by the invention. The gene encoding thedisclosed cDNA is thought to reside on chromosome 19. Accordingly,polynucleotides related to this invention are useful as a marker inlinkage analysis for chromosome 19.

[0079] This gene is primarily expressed in the fetal liver/spleen andpituitary gland.

[0080] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune,hematopoietic, or endocrine disorders, particularly cancers and otherproliferatve conditions. Similarly, polypeptides and antibodies directedto these polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of thehepatic, immune and hematopoetic systems, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. liver, spleen, pituitary,developmental, and cancerous and wounded tissues) or bodily fluids(e.g., lymph, amniotic fluid, serum, plasma, urine, synovial fluid orspinal fluid) or another tissue or cell sample taken from an individualhaving such a disorder, relative to the standard gene expression level,i.e., the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0081] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:55 as residues: Ser-62 to Cys-71, Thr-78 to Leu-86, Ser-104 toLys-109, Ser-130 to Ala-135, Gln-168 to Asp-174.

[0082] The tissue distribution in fetal liver/spleen indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the diagnosis and treatment of hepatic disorders, and disorders ofthe immune and hematopoetic systems, such as hepatic failure, hepatitis,alchoholic liver diseases, portal hypertension, toxic liver injury,liver transplantation, and neoplasm of the liver. The expression in thefetal liver spleen also indicate its function in hematopiesis, andtherefore the gene may be useful in hematopoietic disorders includinganemia, leukemia or cancer radiotherapy/chemotherapy. The expression inthe pituitary land may indicate its use in endocrine disorders withsystemic or specific manifestations. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0083] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:19 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1323 of SEQID NO:19, b is an integer of 15 to 1337, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:19, and whereb is greater than or equal to a+14.

[0084] FEATURES OF PROTEIN ENCODED BY GENE NO: 10

[0085] The translation product of this gene shares sequence homologywith a DNA binding protein from Gallus gallus which is thought to beimportant in transcriptional regulation of gene expression. In specificembodiments, polpeptides of the invention comprise the sequence:MMFGGYETI, (SEQ ID NO:129) YRDESSSELSVDSEVEFQLYSQIH, (SEQ ID NO:130)YAQDLDDVIREEEHEEKNSGNSESSSSKPNQKKL (SEQ ID NO:131)IVLSDSEVIQLSDGSEVITLSDEDSIYRCKGKNV RVQAQENAHGLSSSLQSNELVDKKCKSDIEKPKSEERSGVIREVMIIIEVSSSEEEESTISEGDNVES W, MLLGCEVDDKDDDILLNLVGCENSVTEGEDGINW(SEQ ID NO:132) SIS, DKDIEAQIANNRTPGRWT, (SEQ ID NO:133)QRYYSANKNIICRNCDKRGHLSKNCPLPRKV, (SEQ ID NO:134) and/or RRCFLCSRRGHLLYSCPAPLCEYCPVPKMLDHS (SEQ ID NO:135) CLFRHSWDKQCDRCHMLGHYTDACTEIWRQYHLTTKPGPPKKPKTPSRPSALAYCYHCAQKGHYGHEC PEREVYDPSPVSPFICYYXDKYEIQEREKRLKQKIKVXKKNGVIPEPSKLPYIKAANENPHHDIRKGR ASWKSNRWPQ.

[0086] Polynucleotides encoding these polypeptides are also encompassedby the invention.

[0087] This gene is expressed in tonsils and bone marrow.

[0088] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, disordersof the immune, hematopoetic, and lymphatic systems. Similarly,polypeptides and antibodies directed to these polypeptides are useful inproviding immunological probes for differential identification of thetissue(s) or cell type(s). For a number of disorders of the abovetissues or cells, particularly of the immune, hematopoetic, and lymphsystems, expression of this gene at significantly higher or lower levelsmay be routinely detected in certain tissues or cell types (e.g. immune,hematopoietic, and cancerous and wounded tissues) or bodily fluids(e.g., lymph, serum, plasma, urine, synovial fluid or spinal fluid) oranother tissue or cell sample taken from an individual having such adisorder, relative to the standard gene expression level, i.e., theexpression level in healthy tissue or bodily fluid from an individualnot having the disorder.

[0089] The tissue distribution in bone marrow and tonsil tissues,combined with the homology to a DNA binding protein indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the treatment and diagnosis of disorders in the immune,hematopoetic, and lymph systems. Furthermore, expression of this geneproduct in tonsils indicates a role in the regulation of theproliferation; survival; differentiation; and/or activation ofpotentially all hematopoietic cell lineages, including blood stem cells.This gene product may be involved in the regulation of cytokineproduction, antigen presentation, or other processes that may alsosuggest a usefulness in the treatment of cancer (e.g. by boosting immuneresponses). Since the gene is expressed in cells of lymphoid origin, thegene or protein, as well as, antibodies directed against the protein mayshow utility as a tumor marker and/or immunotherapy targets for theabove listed tissues. Therefore it may be also used as an agent forimmunological disorders including arthritis, asthma, immune deficiencydiseases such as AIDS, leukemia, rheumatoid arthritis, inflammatorybowel disease, sepsis, acne, and psoriasis. In addition, this geneproduct may have commercial utility in the expansion of stem cells andcommitted progenitors of various blood lineages, and in thedifferentiation and/or proliferation of various cell types. Protein, aswell as, antibodies directed against the protein may show utility as atumor marker and/or immunotherapy targets for the above listed tissues.

[0090] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:20 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1376 of SEQID NO:20, b is an integer of 15 to 1390, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:20, and whereb is greater than or equal to a+14.

[0091] FEATURES OF PROTEIN ENCODED BY GENE NO: 11

[0092] One embodiment of this gene comprises polypeptides of thefollowing amino acid sequence: (SEQ ID NO:136)MSFPPHLNRPPMGIPALPPGIPPPQFPGFPPPVPPGTPMIPVPMSIMAPAPTVLVPTVSMVGKHLGARKDHPGLKAKENDENCGPTTTVFVGNISEKASDMLIRQLLAKCGLVLSWKRVQGASGKLQAFGFCEYKEPESTLRALRLLHDLQIGEKKLLVKVDAKTKAQLDEWKAKKKASNGNARPETVTNDDEEALDEETKRRDQMIKGAIEVLIREYSSELNAPSQESDSHPRKKKKEKKEDIFRRFPVAPLIPYPLITKEDINAIEMEEDKRDLISREISKFRDTHKKLEEEKGKKEKERQEIEKERRERERERERERERREREREREREREREKEKERERERERDRDRDRTKERDRDRDRERDRDRDRERSSDRNKDRIRSREKSRDRERERERERE RERERERERERERERE,(SEQ ID NO:137) MSFPPHLNRPPMGIPALPPGIPPPQFPGFPPPVPPGTPMIPVP (SEQ IDNO:138) MSIMAPAPTVLVPTVSMVGKHLGARKDHPGLKAKE, (SEQ ID NO:139) NDENCGPTTTVFVGNISEKASDMLIRQLLAKCGLVLSWKRV, (SEQ ID NO:140) QGASGKLQAFGFCEYKEPESTLRALRLLHDLQIGEKKLLV, (SEQ ID NO:141) KVDAKTKAQLDEWKAKKKASNGNARPETVTNDDEEALDE, (SEQ ID NO:142) ETKRRDQMIKGAIEVLIREYSSELNAPSQESDSHPRKKKK, (SEQ ID NO:143) EKKEDIFRRFPVAPLIPYPLITKEDINAIEMEEDKRDLISREIS, (SEQ ID NO:144) KFRDTHKKLEEEKGKKEKERQEIEKERRERERERERERERR, (SEQ ID NO:145) EREREREREREREKEKERERERERDRDRDRTKERDRDRDRE, and/or (SEQ ID NO:146)RDRDRDRERSSDRNKDRIRSREKSRDRERERERERERERERERERERERE RE.

[0093] Polynucleotides encoding these polypeptides are also encompassedby the invention.

[0094] This gene is expressed in fetal liver/spleen, dendritic, andT-cells.

[0095] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune orhematopoietic disorders. Similarly, polypeptides and antibodies directedto these polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theimmune system, expression of this gene at significantly higher or lowerlevels may be routinely detected in certain tissues or cell types (e.g.immune, hematopoietic, and cancerous and wounded tissues) or bodilyfluids (e.g., lymph, serum, plasma, urine, synovial fluid or spinalfluid) or another tissue or cell sample taken from an individual havingsuch a disorder, relative to the standard gene expression level, i.e.,the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0096] The tissue distribution in immune cells and tissues indicatesthat the protein products of this gene are useful for the treatment anddiagnosis of immune system disorders, particularly those involvingdendritic or T-cells such as inflammation. Furthermore, this geneproduct is primarily expressed in hematopoietic cells and tissues,suggesting that it plays a role in the survival, proliferation, and/ordifferentiation of hematopoieitic lineages. This is particularlysupported by the expression of this gene product in fetal liver, whichis a primary site of definitive hematopoiesis. Expression of this geneproduct in T cells and primary dendritic cells also strongly indicates arole for this protein in immune function and immune surveillance.Protein, as well as, antibodies directed against the protein may showutility as a tumor marker and/or immunotherapy targets for the abovelisted tissues.

[0097] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:21 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1417 of SEQID NO:21, b is an integer of 15 to 1431, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:21, and whereb is greater than or equal to a+14.

[0098] FEATURES OF PROTEIN ENCODED BY GENE NO: 12

[0099] Preferred polypeptides encoded by this gene include thefollowing: (SEQ ID NO:150)MGLNPPGLTSALKPQMEGRLVGGGGSFSSRGRHPAGWVLPQPCLLL SPTLSFPPACGLLVPSPSLLPAVSSYHLPLGRGLIRPAFKIKVCSKLTVWCSL PSPSRWRCCHGNAVALPALGPWRXWEQGSAVRSPAFPVRQAWLPCSGSL TSW, (SEQ ID NO:147)KPQMEGRLVGGGGSFSSRGRHP, (SEQ ID NO:148) LLVPSPSL LPAVSSYHLPLGRGLIR,and/or (SEQ ID NO:149) EQGSAVRSPAFPVR QA WLPCSGS.

[0100] LPAVSSYHLPLGRGLIR (SEQ ID NO: 148), and/or EQGSAVRSPAFPVR QAWLPCSGS (SEQ ID NO: 149).

[0101] Also provided are polynucleotides encoding such polypeptides.

[0102] This gene is expressed in activated neutrophils, endothelialcells, T cells, and to a lesser extent, in brain and liver.

[0103] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune,hematopoietic, integumentary, or vascular disorders, particularlyimmunodeficiencies such as AIDS, and susceptiblity to infectiousdisease. Similarly, polypeptides and antibodies directed to thesepolypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theimmune system and skin, expression of this gene at significantly higheror lower levels may be routinely detected in certain tissues or celltypes (e.g. brain, immune, integumentary, and cancerous and woundedtissues) or bodily fluids (e.g., lymph, serum, plasma, urine, synovialfluid or spinal fluid) or another tissue or cell sample taken from anindividual having such a disorder, relative to the standard geneexpression level, i.e., the expression level in healthy tissue or bodilyfluid from an individual not having the disorder.

[0104] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:58 as residues: Glu-41 to Val-46.

[0105] The tissue distribution in immune cells indicates that the geneproduct is useful for the diagnosis and/or treatment of a variety ofdisorders, including hematopoietic disorders, neurological disorders,liver disease, and disorders involving angiouenesis. The expression ofthis gene product in hematopoietic cells and tissues indicates that itplays a role in the survival, proliferation, and/or differentiation ofhematopoieitic lineages. This is particularly supported by theexpression of this gene product in fetal liver, which is a primary siteof definitive hematopoiesis. Expression of this gene product in T cellsand neutrophils also strongly indicates a role for this protein inimmune function and immune surveillance. Additionally, the tissuedistribution indicates that polynucleotides and polypeptidescorresponding to this gene are useful for the detection/treatment ofneurodegenerative disease states and behavioural disorders such asAlzheimers Disease, Parkinsons Disease, Huntingtons Disease, TouretteSyndrome, schizophrenia, mania, dementia, paranoia, obsessive compulsivedisorder, panic disorder, learning disabilities, ALS, psychoses ,autism, and altered bahaviors, including disorders in feeding, sleeppatterns, balance, and perception. In addition, the gene or gene productmay also play a role in the treatment and/or detection of developmentaldisorders associated with the developing embryo, or sexually-linkeddisorders. Moreover, expression within endothelial cells indicates thatthe protein is useful in the treatment and/or diagnosis of a variety ofvascular disorders, which include, but are not limited to, embolism,aneurysm, microvascular disease, atherosclerosis, and stroke. Protein,as well as, antibodies directed against the protein may show utility asa tumor marker and/or immunotherapy targets for the above listedtissues.

[0106] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:22 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 2525 of SEQID NO:22, b is an integer of 15 to 2539, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:22, and whereb is greater than or equal to a+14.

[0107] FEATURES OF PROTEIN ENCODED BY GENE NO: 13

[0108] This gene is expressed in keratinocytes, and to a lesser extent,in endothelial cells and placenta.

[0109] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,integumentary or vascular disorders, particularly impaired wound healingand autoimmune disorders. Similarly, polypeptides and antibodiesdirected to these polypeptides are useful in providing immunologicalprobes for differential identification of the tissue(s) or cell type(s).For a number of disorders of the above tissues or cells, particularly ofthe skin, expression of this gene at significantly higher or lowerlevels may be routinely detected in certain tissues or cell types (e.g.integumentary, endothelial, and cancerous and wounded tissues) or bodilyfluids (e.g., lymph, serum, plasma, urine, synovial fluid or spinalfluid) or another tissue or cell sample taken from an individual havingsuch a disorder, relative to the standard gene expression level, i.e.,the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0110] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:59 as residues: Pro-35 to Trp-42, Ala-53 to Asp-62, Arg-103 toPro-113.

[0111] The tissue distribution in keratinocytes indicates that theprotein products of this gene are useful for the treatment of woundhealing deficiency and skin disorders, including congenital disorders(i.e. nevi, moles, freckles, Mongolian spots, hemangiomas, port-winesyndrome), integumentary tumors (i.e. keratoses, Bowen's disease, basalcell carcinoma, squamous cell carcinoma, malignant melanoma, Paget'sdisease, mycosis fungoides, and Kaposi's sarcoma), injuries andinflammation of the skin (i.e.wounds, rashes, prickly heat disorder,psoriasis, dermatitis), atherosclerosis, uticaria, eczema,photosensitivity, autoimmune disorders (i.e. lupus erythematosus,vitiligo, dermatomyositis, morphea, scieroderma, pemphigoid, andpemphigus), keloids, striae, erythema, petechiae, purpura, andxanthelasma. Moreover, such disorders may predispose increasedsusceptibility to viral and bacterial infections of the skin (i.e. coldsores, warts, chickenpox, molluscum contagiosum, herpes zoster, boils,cellulitis, erysipelas, impetigo, tinea, althletes foot, and ringworm).Moreover, expression within endothelial cells and pancreatic tissuesindicates that the protein product of this gene may be useful in thetreatment and/or prevention of a variety of vascular disorders,particularly those involving highly vascularized tissues, which include,but are not limited to, embolism, aneurysm, stroke, atherosclerosis, andmiscrovascular disease. Protein, as well as, antibodies directed againstthe protein may show utility as a tumor marker and/or immunotherapytargets for the above listed tissues.

[0112] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:23 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1027 of SEQID NO:23, b is an integer of 15 to 1041, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:23, and whereb is greater than or equal to a+14.

[0113] FEATURES OF PROTEIN ENCODED BY GENE NO: 14

[0114] Preferred polypeptides encoded by this gene comprise thefollowing amino acid sequence: NVTKITLESFLAWKKRKRQEKIDKLEQDMERRKADFKAGKAL VISGREVFEFRPELVNDDDEEA (SEQ ID NO:151), and/orERRKADFKAGKALV ISCREVFE (SEQ ID NO:152). Polynucleotides encoding thesepolypeptides are also encompassed by the invention. The gene encodingthe disclosed cDNA is thought to reside on chromosome 2. Accordingly,polynucleotides related to this invention are useful as a marker inlinkage analysis for chromosome 2.

[0115] This gene is expressed in kidney, and to a lesser extent, inembryonic tissues.

[0116] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, renal,urogenital, or develpomental disorders, particularly renal failure.Similarly, polypeptides and antibodies directed to these polypeptidesare useful in providing immunological probes for differentialidentification of the tissue(s) or cell type(s). For a number ofdisorders of the above tissues or cells, particularly of the kidney,expression of this gene at significantly higher or lower levels may beroutinely detected in certain tissues or cell types (e.g. kidney,developmental, and cancerous and wounded tissues) or bodily fluids(e.g., lymph, amniotic fluid, serum, plasma, urine, synovial fluid orspinal fluid) or another tissue or cell sample taken from an individualhaving such a disorder, relative to the standard gene expression level,i.e., the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0117] The tissue distribution in kidney indicates that this gene orgene product could be used in the treatment and/or detection of kidneydiseases including renal failure, nephritus, renal tubular acidosis,proteinuria, pyuria, edema, pyelonephritis, hydronephritis, nephroticsyndrome, crush syndrome, glomerulonephritis, hematuria, renal colic andkidney stones, in addition to Wilms Tumor Disease, and congenital kidneyabnormalities such as horseshoe kidney, polycystic kidney, and Falconi'ssyndrome. Expression within embryonic tissue indicates that this proteinmay play a role in the regulation of cellular division, and may showutility in the diagnosis and treatment of cancer and other proliferativedisorders. Similarly, developmental tissues rely on decisions involvingcell differentiation and/or apoptosis in pattern formation. Thus thisprotein may also be involved in apoptosis or tissue differentiation andcould again be useful in cancer therapy. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0118] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:24 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1948 of SEQID NO:24, b is an integer of 15 to 1962, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:24, and whereb is greater than or equal to a+14.

[0119] FEATURES OF PROTEIN ENCODED BY GENE NO: 15

[0120] The gene encoding the disclosed cDNA is thought to reside onchromosome 22. Accordingly, polynucleotides related to this inventionare useful as a marker in linkage analysis for chromosome 22.

[0121] This gene is expressed primarily in brain, and to a lesserextent, in liver.

[0122] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, neural orhepatic disorders, particularly depression, manic depression and otherneurodegenerative diseases. Similarly, polypeptides and antibodiesdirected to these polypeptides are useful in providing immunologicalprobes for differential identification of the tissue(s) or cell type(s).For a number of disorders of the above tissues or cells, particularly ofthe central nervous system, expression of this gene at significantlyhigher or lower levels may be routinely detected in certain tissues orcell types (e.g. brain, liver, cancerous and wounded tissues) or bodilyfluids (e.g., lymph, bile, serum, plasma, urine, synovial fluid orspinal fluid) or another tissue or cell sample taken from an individualhaving such a disorder, relative to the standard gene expression level,i.e., the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0123] The tissue distribution in brain indicates that the proteinproducts of this gene are useful for the treatment of central nervoussystem disorders such as depression and other mental illnesses, andneurodegenerative disorders, such as Alzheimers Disease, ParkinsonsDisease, Huntingtons Disease, Tourette Syndrome, schizophrenia, mania,dementia, paranoia, obsessive compulsive disorder, panic disorder,learning disabilities, ALS, psychoses , autism, and altered bahaviors,including disorders in feeding, sleep patterns, balance, and perception.In addition, the gene or gene product may also play a role in thetreatment and/or detection of developmental disorders associated withthe developing embryo, or sexually-linked disorders. Moreover, theexpression within liver tissue indicates that polynucleotides andpolypeptides corresponding to this gene are useful for the detection andtreatment of liver disorders and cancers (e.g. hepatoblastoma, jaundice,hepatitis, liver metabolic diseases and conditions that are attributableto the differentiation of hepatocyte progenitor cells). Protein, as wellas, antibodies directed against the protein may show utility as a tumormarker and/or immunotherapy targets for the above listed tissues.

[0124] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:25 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1214 of SEQID NO:25, b is an integer of 15 to 1228, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:25, and whereb is greater than or equal to a+14.

[0125] FEATURES OF PROTEIN ENCODED BY GENE NO: 16

[0126] The translation product of this gene shares homology withPrefoldin, which is believed to function as a chaperone that deliversunfolded proteins to cytosolic chaperonins (Cell 93, 863-873 (1998)). Inspecific embodiments, polypeptides of the invention comprise thefollowing amino acid sequence: (SEQ ID NO:153) MCDELPGE GRWEPGQDRKLCLSFPLGTPARPIKSVCPTLLSLVFLSRGMEQRVREA VA VSTSAPA PSASEPFLSWGMGLAXFSFPFLYL, (SEQ ID NO:154) GASLGSSSSCPSH SWWGQRSVCRETASPLPRWMLYLDGLATSHFLHHPEPHLLPSPGVFTRLCCHLCPGHXS LSGCVMNSQEREDGSQGKIGSSA, (SEQ ID NO:155) TSVLS SSSVYCMQARKLSVSQRYRKGKEKXARPIPQERKGSDAEGAGAEVETATASLTLCSIPLLKKTRL SRVGQTLFIGLAGVPSGKLRQSFLSCPGSHLPSPGSSSHIPRGKXVLGR GGSKAG, (SEQ ID NO:156)ALVKGTGREKRRXQGPSPKKGRALMQREQELRWRRPLm PLSPSVPSLCSRKPGLAEWDRRFLLVWLACLVESSGRASYLALAPIFPL LGVHHTSREGXVSWAEVAAKPGKNSRAGKQMGLRVMQKM,(SEQ ID NO:157) SFPLGTPARPIKSVCPTLLSLVFLSRGMEQRV, (SEQ ID NO:158)TASPLPRWM LYLDGLATSHFLHHPEPHLLPS, (SEQ ID NO:159) RKGSDAEGAGAEVETATASLTLCSIPLLKKT, (SEQ ID NO:160) QREQELRWRRPLPLSPSVPSLCSRK, and/or (SEQID NO:161) PLLGVHHTSREGXVSWAEVAAKPGKNSRA.

[0127] Polynucleotides encoding these polypeptides are also encompassedby the invention. The gene encoding the disclosed cDNA is thought toreside on chromosome 5. Accordingly, polynucleotides related to thisinvention are useful as a marker in linkage analysis for chromosome 5.

[0128] This gene is expressed in fetal brain, and to a lesser extent, inplacenta, endothelial cells, fetal lung, and T cells.

[0129] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, neural ordevelopmental disorders, particularly restinosis, birth defects andimmune disorders. Similarly, polypeptides and antibodies directed tothese polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of thecardiovascular system, and developmental process, expression of thisgene at significantly higher or lower levels may be routinely detectedin certain tissues or cell types (e.g. brain, endothelial cells,develpomental, immune, pulmonary, and cancerous and wounded tissues) orbodily fluids (e.g., lymph, pulmonary surfactant or sputum, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0130] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:62 as residues: Gln-36 to Lys-42, Glu-89 to Arg-104.

[0131] The tissue distribution in brain, combined with the homology toprefoldin indicates that the protein products of this gene are usefulfor the development of agonists and/or antagonists for for treatment ofnervous system disorders and fetal developmental disorder, and indicatesthat the translation product of this gene is useful in the detectionand/or treatment of diseases associated with the improper delivery ofunfolded proteins to cytosolic chaperoning. Furthermore, the tissuedistribution indicates that polynucleotides and polypeptidescorresponding to this gene are useful for the detection/treatment ofneurodegenerative disease states and behavioural disorders such asAlzheimers Disease, Parkinsons Disease, Huntingtons Disease, TouretteSyndrome, schizophrenia, mania, dementia, paranoia, obsessive compulsivedisorder, panic disorder, learning disabilities, ALS, psychoses ,autism, and altered bahaviors, including disorders in feeding, sleeppatterns, balance, and perception. In addition, the gene or gene productmay also play a role in the treatment and/or detection of developmentaldisorders associated with the developing embryo, sexually-linkeddisorders, or disorders of the cardiovascular system. Protein, as wellas, antibodies directed against the protein may show utility as a tumormarker and/or immunotherapy targets for the above listed tissues.

[0132] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:26 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1326 of SEQID NO:26, b is an integer of 15 to 1340, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:26, and whereb is greater than or equal to a+14.

[0133] FEATURES OF PROTEIN ENCODED BY GENE NO: 17

[0134] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: MSVLKGERQQTLALAVLSVAKENA RDVCCLQGWQDTSCRDTSCAALRGGLQTLFPAPVHFRCGGPAELKGRGS (SEQ ID NO:162),AHSFTTPEEARGAGSMGCRFPFKHTHSPHPRRPEVQGAWAGCTSAGEKAEPP PSREPGSQAS RFPLPP(SEQ ID NO:163), GWQDTSCRDTSCAALRGGLQTLFPA (SEQ ID NO:164), and/orGCRFPFKHTHSPHPRRPEVQGAWA (SEQ ID NO:165). Polynucleotides encoding thesepolypeptides are also encompassed by the invention.

[0135] This gene is expressed in hemangiopericytoma, and to a lesserextent, in fetal tissues.

[0136] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,developmental disorders, particularly hemangiopericytomas and othersoft-tissue cancers, as well as other proliferative disorders.Similarly, polypeptides and antibodies directed to these polypeptidesare useful in providing immunological probes for differentialidentification of the tissue(s) or cell type(s). For a number ofdisorders of the above tissues or cells, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. fetal, kidney, and cancerous andwounded tissues) or bodily fluids (e.g., lymph, amniotic fluid, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0137] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:63 as residues: Glu-43 to Pro-51, Gly-71 to Ar-82, Pro-96 toArg-103, Thr-130 to Gly-140.

[0138] The polynucleotides and polypeptides related to this gene arebelieved to be useful for the treatment and diagnosis of tumors,particularly hemangiopericytomas, and for the treatment of developmentaldisorders. Furthermore, the tissue distribution indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the diagnosis and/or treatment of disorders of the developingembryo, where it may play a crucial role in the development and/orsurvival of the developing embryo or fetus. Expression of this geneproduct in a vascular-rich tissue such as the placenta also indicatesthat this gene product may be produced more generally in endothelialcells or within the circulation. In such instances, it may play moregeneralized roles in vascular function, such as in angiogenesis. It mayalso be produced in the vasculature and have effects on other cellswithin the circulation, such as hematopoietic cells. It may serve topromote the proliferation, survival, activation, and/or differentiationof hematopoietic cells, as well as other cells throughout the body.

[0139] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:27 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 792 of SEQID NO:27, b is an integer of 15 to 806, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:27, and where bis greater than or equal to a+14.

[0140] FEATURES OF PROTEIN ENCODED BY GENE NO: 18

[0141] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: (SEQ ID NO:166)PHQVEGRLGTMETWDSSHEGLLHCRIPLKGSWVQEPSCQYQWRRTRCMGI PPATSGWPCRAPAFLCARAEFPASPGGSTNF, (SEQ ID NO:167) L VTPPSGGETGDHGNMGQLPRRALALQNSTQGILGPGAELPVSVEKDKVHGDPASNIRMA MPGTRFPLCSCRIPCQPGGIH, (SEQ ID NO:168) EGLLHCRIPLKGSWVQEPS CQYQWRRTRCMGI, and/or(SEQ ID NO:169) QNSTQGILGPGAELPVSVEKDK VHGDPAS.

[0142] Polynucleotides encoding these polypeptides are also encompassedby the invention.

[0143] This gene is expressed in fetal liver, and to a lesser extent, inbrain and T cells.

[0144] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, neural,developmental, hepatic, or immune and hematopoietic disorders.Similarly, polypeptides and antibodies directed to these polypeptidesare useful in providing immunological probes for differentialidentification of the tissue(s) or cell type(s). For a number ofdisorders of the above tissues or cells, particularly of the hepaticsystem, nervous system and immune system, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. immune, liver, brain, cancerous andwounded tissues) or bodily fluids (e.g., lymph, amniotic fluid, bile,serum, plasma, urine, synovial fluid or spinal fluid) or another tissueor cell sample taken from an individual having such a disorder, relativeto the standard gene expression level, i.e., the expression level inhealthy tissue or bodily fluid from an individual not having thedisorder.

[0145] The tissue distribution in brain indicates that the proteinproducts of this gene are useful for the identification of agonistsand/or antagonists for the treatment of mental illnesses such asschizophrenia and depression. The gene product may also be useful formonitoring fetal development during pregnancy. Furthermore, theexpression of this gene product in fetal liver indicates a role in theregulation of the proliferation; survival; differentiation; and/oractivation of potentially all hematopoietic cell lineages, includingblood stem cells. This gene product may be involved in the regulation ofcytokine production, antigen presentation, or other processes that mayalso suggest a usefulness in the treatment of cancer (e.g. by boostingimmune responses). Since the gene is expressed in cells of lymphoidorigin, the gene or protein, as well as, antibodies directed against theprotein may show utility as a tumor marker and/or immunotherapy targetsfor the above listed tissues. Therefore it may be also used as an agentfor immunological disorders including arthritis, asthma, immunedeficiency diseases such as AIDS, leukemia, rheumatoid arthritis,inflammatory bowel disease, sepsis, acne, and psoriasis. In addition,this gene product may have commercial utility in the expansion of stemcells and committed progenitors of various blood lineages, and in thedifferentiation and/or proliferation of various cell types.Alternatively, the tissue distribution indicates that polynucleotidesand polypeptides corresponding to this gene are useful for the detectionand treatment of liver disorders and cancers (e.g. hepatoblastoma,jaundice, hepatitis, liver metabolic diseases and conditions that areattributable to the differentiation of hepatocyte progenitor cells).Protein, as well as, antibodies directed against the protein may showutility as a tumor marker and/or immunotherapy targets for the abovelisted tissues.

[0146] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:28 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly. preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 682 of SEQID NO:28, b is an integer of 15 to 696, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:28, and where bis greater than or equal to a+14.

[0147] FEATURES OF PROTEIN ENCODED BY GENE NO: 19

[0148] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: (SEQ ID NO:170)FGTRKKYHLCMIPNLDLNLDRDLVLPDVSYQVESSEEDQSQT, (SEQ ID NO:171)FLLSLGSLVMLLQDLVHSELDGTLHYTVALHKDGIEMSCEQSIDSPDFHL LDWKCTVEIHKEKKQQSLSLRIHSLRLILLTGFHLITXIWKHQISIQIE IQIGY HTQMVFFPRAE, (SEQ IDNO:172) VHSELDGTLHYTVALHKDGIEMSCEQ, and/or (SEQ ID NO:173)QSLSLRIHSLRLILLTGFHLITXIWKIHQ.

[0149] Polynucleotides encoding these polypeptides are also encompassedby the invention.

[0150] This gene is expressed in T cells, fetal liver, and to a lesserextent in brain.

[0151] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, centralnervous diseases and immune or hematopoietic disorders. Similarly,polypeptides and antibodies directed to these polypeptides are useful inproviding immunological probes for differential identification of thetissue(s) or cell type(s). For a number of disorders of the abovetissues or cells, particularly of the central nervous system and immunesystem, expression of this gene at significantly higher or lower levelsmay be routinely detected in certain tissues or cell types (e.g. liver,immune, brain, cancerous and wounded tissues) or bodily fluids (e.g.lymph, serum, bile, amniotic fluid, plasma, urine, synovial fluid orspinal fluid) or another tissue or cell sample taken from an individualhaving such a disorder, relative to the standard gene expression level,i.e., the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0152] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:65 as residues: Lys-69 to Leu-74, Ser-92 to Phe-97, Asp-109 toLeu-117, Leu-142 to Ser-159, Thr-166 to Glu-183, Ala-191 to Glu-205,Pro-213 to Glu-220.

[0153] The tissue distribution in T-cells and brain indicates that theprotein products of this gene are useful for the development of drugsfor the treatment of disorders affecting the central nervous system andimmune system. The expression of this gene product primarily inhematopoietic cells and tissues indicates that it plays a role in thesurvival, proliferation, and/or differentiation of hematopoieiticlineages. This is particularly supported by the expression of this geneproduct in fetal liver, which is a primary site of definitivehematopoiesis. Expression of this gene product in T cells also stronglyindicates a role for this protein in immune function and immunesurveillance. Protein, as well as, antibodies directed against theprotein may show utility as a tumor marker and/or immunotherapy targetsfor the above listed tissues.

[0154] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:29 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore, polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 993 of SEQID NO:29, b is an integer of 15 to 1007, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:29, and whereb is greater than or equal to a+14.

[0155] FEATURES OF PROTEIN ENCODED BY GENE NO: 20

[0156] The translation product of this gene shares sequence homologywith a C.elegans ORF that seems to be a transmembrane protein. (SeeAccession No. 790406.) In specific embodiments, polypeptides of theinvention comprise the following amino acid sequence: (SEQ ID NO:174)MAAACGPGAAGTACSSACICFCDRGPCLGWNDPDRMLLRDVKALTLHYDR YTTSRSWIPSHSPQLKCVGGTAGCDSYTPKVIQCQNKGWDGYDVQWECK TDLDIAYKFGKTVVSCEGYESSEDQYVLRGSCGLEYNLDYTELGLQKLK ESGKQHGFASFSDYYYKWSSADSCNMSGLITIVVLLGIAFVVYKLFLSD GQYSPPPYSEYPPFSHRYQRFTNSAGPPPPGFKSEFTGPQNTG HGATSGFGSAFTGQQGYENSGPGFWTGLGTGGILGYLFGSNRAATPFSDSWYYPSY PPSYPGTWNRAYSPLHGGSGSYSVCSNSDTKTRTASGYGGTRRR, (SEQ ID NO:175)ACSSACICFCDRGPCLGWNDPDRM, (SEQ ID NO:176) TAGCDSYTPKV IQCQNKGWDGYDVQW,(SEQ ID NO:177) EYNLDYTELGLQKLKESGK QHGFASFSDYYYK, (SEQ ID NO:178)YKLFLSDGQYSPPPYSEYPPFSHRYQRF, (SEQ ID NO:179)ENSGPGFWTGLGTGGILGYLFGSNRA, and/or (SEQ ID NO:180)NRAYSPLHGGSGSYSVCSNSDTKTR.

[0157] Polynucleotides encoding these polypeptides are also encompassedby the invention.This gene maps to chromosome 8, and therefore can beused as a marker in linkage analysis for chromosome 8.

[0158] This gene is strongly expressed in fetal liver/spleen andpancreas, and to a lesser extent in T-cells, primary dendritic cells,bone marrow cells, and amygdala.

[0159] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,reproductive, immune, hematopoietic, neural, or endocrine disorders,particularly cancer. Similarly, polypeptides and antibodies directed tothese polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theimmune system and endocrine system, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. immune, pancreas, brain, cancerousand wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,urine, synovial fluid or spinal fluid) or another tissue or cell sampletaken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0160] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:66 as residues: Gly-30 to Asp-35, Tyr49 to Asp-59, Ala-71 toThr-77, Gln-84 to Asp-92, Gln-94 to Thr-99, Glu-115 to Val-125, Lys-145to Lys-151, Ser-159 to Asn-172, Asp-196 to Phe-228, Ser-230 to His-240,Gln-253 to Gly-262, Pro-285 to Tyr-291, Pro-293 to Trp-303, Leu-310 toSer-316, Ser-321 to Arg-339.

[0161] The tissue distribution combined with the homology to a C.elegans transmembrane-like protein indicates that the protein product ofthis gene plays a role important in both vertebrates and invertegratesand is useful for the diagnosis or treatment of disorders related tothis gene. Furthermore, the tissue distribution indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the diagnosis and/or treatment of hematopoietic disorders. This geneproduct is primarily expressed in hematopoietic cells and tissues,suggesting that it plays a role in the survival, proliferation, and/ordifferentiation of hematopoieitic lineages. This is particularlysupported by the expression of this gene product in fetal liver and bonemarrow, the two primary sites of definitive hematopoiesis. Expression ofthis gene product in T cells and primary dendritic cells also stronglyindicates a role for this protein in immune function and immunesurveillance. Alternatively, the tissue distribution indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the detection, treatment, and/or prevention of various endocrinedisorders and cancers, particularly Addison's disease, Cushing'sSyndrome, and disorders and/or cancers of the pancrease (e.g. diabetesmellitus), adrenal cortex, ovaries, pituitary (e.g., hyper-,hypopituitarism), thyroid (e.g. hyper-, hypothyroidism), parathyroid(e.g. hyper-,hypoparathyroidism), hypothallamus, and testes. Protein, aswell as, antibodies directed against the protein may show utility as atumor marker and/or immunotherapy targets for the above listed tissues.

[0162] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:30 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 2012 of SEQID NO:30, b is an integer of 15 to 2026, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:30, and whereb is greater than or equal to a+14.

[0163] FEATURES OF PROTEIN ENCODED BY GENE NO: 21

[0164] This gene is expressed primarily in embryonic tissue, testes, andto a lesser extent in ovary, hepatoma, kidney, endothelial, and smoothmuscle cells.

[0165] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, metabolicdisorder, abnormal embryonic development, or reproductive disorders,such as tumors. Similarly, polypeptides and antibodies directed to thesepolypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theembryonic or vascular tissues, expression of this gene at significantlyhigher or lower levels may be routinely detected in certain tissues orcell types (e.g., reproductive, developmental, endothelial, hepatic,renal, and cancerous and wounded tissues) or bodily fluids (e.g., lymph,amniotic fluid, bile, seminal fluid, serum, plasma, urine, synovialfluid or spinal fluid) or another tissue or cell sample taken from anindividual having such a disorder, relative to the standard geneexpression level, i.e., the expression level in healthy tissue or bodilyfluid from an individual not having the disorder.

[0166] The tissue distribution in embryonic and liver tissues, combinedwith the homology to a conserved NADH dehydrogenase indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the diagnosis or treatment of metabolic disorders of embryonicand/or vascular tissues. Furthermore, polynucleotides and polypeptidescorresponding to this gene are useful for the diagnosis and/or treatmentof disorders of the placenta. Specific expression within the testes andovaries indicates that this gene product may play a role in the properestablishment and maintanence of normal ovarian and testicular function.Alternately, this gene product may be produced by the placenta and thentransported to the embryo, where it may play a crucial role in thedevelopment and/or survival of the developing embryo or fetus.Expression of this gene product in a vascular-rich tissue such as theplacenta also indicates that this gene product may be produced moregenerally in endothelial cells or within the circulation. In suchinstances, it may play more generalized roles in vascular function, suchas in angiogenesis. It may also be produced in the vasculature and haveeffects on other cells within the circulation, such as hematopoieticcells. It may serve to promote the proliferation, survival, activation,and/or differentiation of hematopoietic cells, as well as other cellsthroughout the body. Protein, as well as, antibodies directed againstthe protein may show utility as a tumor marker and/or immunotherapytargets for the above listed tissues.

[0167] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:31 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleoctides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 685 of SEQID NO:31, b is an integer of 15 to 699, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:31, and where bis greater than or equal to a+14.

[0168] FEATURES OF PROTEIN ENCODED BY GENE NO: 22

[0169] The translation product of this gene shares sequence homologywith alpha 1C adrenergic receptor which is thought to be important inneuronal signal transmission. In specific embodiments, polypeptides ofthe invention comprise the following amino acid sequence: (SEQ IDNO:181) TESQMKCFLGNSHDTAPRHTCSGQGLHGGXXXTAPLRALQQHSQDGKLCTNSLPAARGGPHKHVVVTVVY SVKHWKPTERSSVSIKKEEETDWDMDQLSKQRTTYEMKSGSSGVQTEELRHPSL, (SEQ ID NO:182) NASWEIHMTQRHVIPXLARASMXVXXXQRPSELCSSIRRMANSAQIVFPLPVGAPTNTLSSLLYT V LNTGNQQKEAV, (SEQ IDNO:183) APLRALQQHSQDGKLCTNSLPAARGGPHKH, (SEQ ID NO:184)RSSVSIKKEEETDWDMDQLSKQRTTYE, and/or (SEQ ID NO: 185)LCSSIRRMANSAQIVFPLPVGAPTNTLSS.

[0170] Polynucleotides encoding these polypeptides are also encompassedby the invention.

[0171] This gene is expressed primarily in breast lymph node, testiculartumors, and to a lesser extent, in uterine cancer.

[0172] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune,reproductive, neural, or proliferative disorders, such as cancers.Similarly, polypeptides and antibodies directed to these polypeptidesare useful in providing immunological probes for differentialidentification of the tissue(s) or cell type(s). For a number ofdisorders of the above tissues or cells, particularly of the neurologic,breast lymph node, uterine cancer, and testicular cancer, expression ofthis gene at significantly higher or lower levels may be routinelydetected in certain tissues or cell types (e.g. breast, testes,cancerous and wounded tissues) or bodily fluids (e.g., lymph, seminalfluid, serum, plasma, urine, synovial fluid or spinal fluid) or anothertissue or cell sample taken from an individual having such a disorder,relative to the standard gene expression level, i.e., the expressionlevel in healthy tissue or bodily fluid from an individual not havingthe disorder.

[0173] The tissue distribution in lymph, testicular, and uterine tissuesindicates that the protein product of this gene may be beneficial in thetreatment and/or prevention of a variety of reproductive disorders, andmay even show utility in ameliorating disorders related to aberrantimmune responses to normal or proliferative tissues. The homology toalpha 1C adrenergic receptor indicates that polynucleotides andpolypeptides corresponding to this gene are useful for transmittingsignals to neurons. Furthermore, the tissue distribution indicates thatthe protein product of this gene is useful for the treatment anddiagnosis of conditions concerning proper testicular function (e.g.endocrine function, sperm maturation), as well as cancer. Therefore,this gene product is useful in the treatment of male infertility and/orimpotence. This gene product is also useful in assays designed toidentify binding agents as such agents (antagonists) are useful as malecontraceptive agents. Similarly, the protein is believed to by useful inthe treatment and/or diagnosis of testicular cancer. The testes are alsoa site of active gene expression of transcripts that may be expressed,particularly at low levels, in other tissues of the body. Therefore,this gene product may be expressed in other specific tissues or organswhere it may play related functional roles in other processes, such ashematopoiesis, inflammation, bone formation, and kidney function, toname a few possible target indications. Protein, as well as, antibodiesdirected against the protein may show utility as a tissue-specificmarker and/or immunotherapy target for the above listed tissues.

[0174] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:32 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1250 of SEQID NO:32, b is an integer of 15 to 1264, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:32, and whereb is greater than or equal to a+14.

[0175] FEATURES OF PROTEIN ENCODED BY GENE NO: 23

[0176] The translation product of this gene shares sequence homologywith G-protein-coupled receptor which is thought to be important inmediating a wide variety of physiological function and belongs to a genesuperfamily with members ranging from chemokine receptor to bradykininreceptor. This gene has also recently been gened by another group,calling the gene platelet activating receptor homolog. (See AccessionNo. 2580588.) Preferred polypeptide fragments comprise the amino acidsequence: LSIIFLAFVSIDRCLQL (SEQ ID NO:186), GSCFATWAFQKNTNHRCVSIYLINLLTADFLLTLALPVKIVVDLGVAPWKLKIFHCQVTACLIYIN (SEQ ID NO:187), and/orKNTNHRCVSIYLINLLTADFLLTLALPVKIV (SEQ ID NO:188). Also preferred arepolynucleotide fragments encoding these polypeptide fragments.

[0177] Chemokines, are soluble, low molecular weight members of thecytokine family that have chemo-attractant activity. Chemokinesselectively induce chemotaxis of the white blood cells, includingleukocytes, such as monocytes, macrophages, eosinophils, basophils, mastcells, and lymphocytes, such as T-cells, B-cells; and polymorphonuclearleukocytes (neutrophils). In addition, Chemokines can selectively induceother changes in responsive cells, including changes in cell shape,transient rises in the concentration of intracellular free calcium([Ca2+]i), granule exocytosis, integrin upregulation, formation ofbioactive lipids (e.g. leukotrienes) and respiratory burst, associatedwith leukocyte activation. Therefore, the chemokines are earlyactivators of the inflammatory response, causing inflammatory mediatorrelease, chemotaxis and extravasation to sites of infection orinflammation.

[0178] The chemokines are related in their primary DNA structure. Theyshare four conserved cysteines, which form disulphide bonds. cDNAcloning and biochemical characterization of several chemokines hasrevealed they have a leader sequence of 20-25 amino acids, that iscleaved upon secretion to yield a protein of approximately 92 to 99amino acids (8 to 10 Kd). The family is divided into two branches basedupon the conserved cysteine motif, designated as the C-C chemolcines (bchemokines) and the C-X-C chemokines (a chemokines), in which the firsttwo conserved cysteines are adjacent, or are separated by, anintervening residue, respectively (Baggiolini, M. and C. A. Dahinden,Immunology Today, 15:127-13 3 (1994)).

[0179] The C-X-C chemokines are potent chemoattractants and activatorsof neutrophils, such as interleukin 8 (IL-8), PF4 andneutrophil-activating peptide 2 (NAP-2). The C-C chemokines includemolecules such as human monocyte chemotactic proteins 1-3 (MCP-1, MCP-2,and NCP-3), RANTES (Regulated on Activation, Normal T Expressed andSecreted), and the macrophage inflammatory proteins 1a and 1b (MIP-1aand MIP-1b). The C-C chemokines have been characterized aschemoattractants and activators of monocytes or lymphocytes, but do notappear to be chemoattractants for neutrophils. For example, recombinantRANTES is a chemo-attractant for monocytes, as well as, for memoryT-cells in vitro (Schall, T. J. et al., Nature, 347:669-671 (1990)).Furthermore, a chemokine called lymphotactin, with a single cysteinepair in the molecule, has been identified that attracts lymphocytes(Kelner, G. S., et al., Science, 266:1395-1399 (1994)).

[0180] The C-C chemokines are of great scientific and pharmaceuticalinterest because of their potential role in allergic inflammation. Forexample, MCP-1 induces exocytosis of human basophils, resulting inrelease of high levels of histamine and leukotriene C4, in addition toother inflammatory mediators. Since these cellular events are triggeredin response to chemokine binding, there remains great interest in thereceptors for the C-C chemokines. Recently, a receptor for C-Cchemokines has been gened and is reported to bind MIP-1a and RANTES.Accordingly, this MIP-1a/RANTES receptor was designated C-C chemokinereceptor 1 (CHR-1; Neote, K. et al., Cell, 72:415-425 (1993); Horuk, R.et al., WO 94/11504, published May 26, 1994; Gao, J. -I. et al. J. Exp.Med., 177: 1421-1427 (1993)). An MCP-1 receptor has also been gened(Charo, I. F. et al., Proc. Natl. Acad. Sci. USA, 91:2752 (1994)). Thisreceptor, designated CKR-2, is reported to bind MCP-1 with high affinityand MCP-3 with lower affinity (ChEO, I, F., et al., Proc. Natl. Acad.Sci. USA, 91:2752-2756 (1994)). CKR-2 has been shown to exist in twoisoforms: Isoform A and B. This dicotomy results from the use of analternative splice site in isoform A mRNA producing a distinctcytoplasmic tail. Isoform B, which is not spliced in this region, hasbeen shown to be a functional receptor for MCP-1 and MCP-3 in bothbinding and signal transduction assays (Charo, I. F. et al., Proc. Natl.Acad. Sci. USA, 91:2752-2756 (1994); Myers, S. J., et al., J. Biol.Chem., 270:5786-5792 (1995)). More recently, a new receptor called CKR-4has been described; cDNA from this receptor was reported to produce aCa2+ activated chloride current in response to MCP-1, MIP-1a and RANTESwhen injected into X. laevis oocytes (Poer, C. A., et al., J. Biol.Chem., 270: 19495-19500 (1995)). Due to the importance of Ca2+ releasein activation of signal transduction pathways in immune cells, oneexperienced in the art would appreciate the role of this receptor as theprimary initiation signal for activating subsequent signalling events.

[0181] Similarly, based upon their DNA structure, the MCP-1 receptor(CKR-2) and C-C chemokine receptor 1 are predicted to belong to asuperfamily of seven transmembrane spanning G-protein coupled receptors(Gerard C., and Gerard N. P., Annu. Rev. Immunol., 12: 775-808 (1994);Gerard C., and Gerard N. P., Curr. Opin. Immnunol., 6:140-145 (1994)).This family of G-protein coupled (serpentine) receptors comprises alarge group of integral membrane proteins containing seventransmembrane-spanning regions. These regions, or domains, are believedto represent conserved transmembrane a-helices comprising 20 to 30hydrophobic amino acids connected by at least eight, divergentextracellular or cytoplasmic loops of primarily hydrophilic amino acids.Most G-protein coupled receptors have single conserved cysteine residueson each of the first two extracellular loops which form disulfide loopsthat are believed to stabilize functional protein structure. The seventransmembrane regions are designated as TM1, TM2, TM3, TM4, TM5, TM6,and TM7. TM3 is also implicated in signal transduction (Probst, W. C.,et al., DNA and Cell Biology, 11:1-20).

[0182] The ligands of these receptors include a diverse group ofmolecules, including small biogenic amine molecules, such as epinethrineand norepinephrine, peptides, such as substance P and neurokinins, andlarger proteins, such as chemokines. The receptors are coupled to Gproteins, which are heterotrimeric regulatory proteins capable ofbinding GTP and mediating signal transduction from coupled receptors.For example, by the production of intracellular mediators. The ligandbinding sites of G-protein coupled receptors are believed to comprise ahydrophilic socket formed by several G-protein receptor transmembranedomains. The hydrophilic side of each G-protein coupled receptortransmembrane helix is postulated to face inward and form the polarligand binding site. TM3 has been implicated in several G-proteincoupled receptors as gaving a ligand binding site, which includes theinvolvement of the TM3 aspartate residue. Additionally, TM5 serines, aTM6 asparagine, and TM6 or TM7 phenylalanines or tyrosines are alsoimplicated in ligand binding.

[0183] It is well established that many medically significant biologicalprocesses are mediated by proteins participating in signal transductionpathways that involve G-proteins and/or second messengers, e.g. cAMP(Lefkowitz, Nature, 351:353-354 (1991)). For example, in one form ofsignal transduction, the effect of hormone binding is activation of anenzyme, adenylate cyclase, inside the cell. Enzyme activation byhormones is dependent upon the presence of the nucleotide GTP, whichitself, has been shown to influence hormone binding. A G-proteinphysically connects the hormone receptors to adenylate cyclase. Uponactivation of the hormone receptor via binding of its receptive ligand,the G-protein was shown to exchange GTP for bound GDP. The GTP-carrying,active form of the G-protein, then binds to an activated adenylatecyclase. Hydrolysis of GTP to GDP, catalyzed by the G-protein itself,returns the G-protein to its basal, inactive form. Thus, the G-proteinserves a dual role, as an intermediate that relays the signal fromreceptor to downstream effector, as well as a clock that controls theduration of the signal.

[0184] G-protein coupled receptors can be coupled intracellularly byheterotrimeric G-proteins to various intracellular enzymes, ionchannels, and transporters (Johnson, et al., Endoc., Rev., 10:317-331(1989)). Different G-protein a-subunits preferentially stimulateparticular effectors to modulate various biological functions in a cell.Phosphorylation of cytoplasmic residues of G-protein coupled receptorshave been identified as an important mechanism for the regulation ofG-protein coupling of some G-protein coupled receptors. G-proteincoupled receptors are found in numerous sites within a mammalian host,including dopamine receptors which bind to neuroleptic drugs used fortreating psychotic and neurological disorders. Other examples of membersof this family include calcitonin, adrenergic, endothelin, cAMP,adenosinse, muscarinic, acetylcholine, serotinin, histamine, thrombin,kinin, follicle stimulating hormone, opsins, endothelial differentiationgene-l receptor, and rhodopsins, odarant, cytomegalovirus receptors,etc.

[0185] Sequencing of two recently gened IL-8 receptor cDNAs haverevealed that these C-X-C receptor proteins also share sequencesimilarity with seven transmembrane-spanning G protein-coupted receptorproteins (Murphy P. M. and H. L. Tiffany, Science, 253:1280-1283 (1991);Murphy et al., WO 93/06299; Holmes, W. E. et al., Science, 253:1278-1290(1991)). Additional receptors for chemotactic proteins such asanaphylatoxin C5a, and bacterial formylated tripeptide fMLP, have beencharacterized by cloning and found to encode receptor proteins whichshare sequence similarity to these transmembrane-spanning proteins(Gerard, N. P. and C. Gerard, Nature, 349: 614-617 (1991); Boulay, F. etal., Biochemistry, 29:11123-11133 (1990)). Although a number of otherproteins and genes have been identified that share significantsimilarity in sequence, tissue expression, and leukocyte subpopulationdistribution, to known chemokine receptors, the ligands for thesereceptors remain undefined. Thus, there is a need to identify otherchemokine receptor polypeptides, as these polypeptides may modulate the.Disturbances of such regulation may be involved in disorders relating tothe immune or hematopoietic system. Therefore, there is a need foridentification and characterization of such human polypeptides which canplay a role in detecting, preventing, ameliorating or correcting suchdisorders. In an attempt to express embodiments of invention via genetherapy, the following protocol may be followed. Fibroblasts areobtained from a subject by skin biopsy. The resulting tissue is placedin tissue-culture medium and separated into small pieces. Small chunksof the tissue are placed on a wet surface of a tissue culture flask,approximately ten pieces are placed in each flask. The flask is turnedupside down, closed tight and left at room temperature over night. After24 hours at room temperature, the flask is inverted and the chunks oftissue remain fixed to the bottom of the flask and fresh media (e.g.,Ham's F12 media, with 10% FBS, penicillin and streptomycin, is added.This is then incubated at 37° C. for approximately one week. At thistime, fresh media is added and subsequently changed every several days.After an additional two weeks in culture, a monolayer of fibroblastsemerge. The monolayer is trypsinized and scaled into larger flasks.pMV-7 (Kirschmeier, P. T. et al, DNA, 7:219-25 (1988) flanked by thelong terminal repeats of the Moloney murine sarcoma virus, is digestedwith EcoRI and HindIII and subsequently treated with calf intestinalphosphatase.

[0186] The linear vector is fractionated on agarose gel and purified,using glass beads. The cDNA encoding a polypeptide of the presentinvention is amplified using PCR primers which correspond to the 5′ and3′ end sequences respectively. The 5′ primer contains an EcoRI site andthe 3′ primer contains a HindIII site. Equal quantities of the Moloneymurine sarcoma virus linear backbone and the EcoRI and HindIII fragmentare added together, in the presence of T4 DNA ligase. The resultingmixture is maintained under conditions appropriate for ligation of thetwo fragments. The ligation mixture is used to transform bacteria HB101,which are then plated onto agar-containing kanamycin for the purpose ofconfirming that the vector had the gene of interest properly inserted.The amphotropic pA317 or GP+am12 packaging cells are grown in tissueculture to confluent density in Dulbecco's Modified Eagles Medium (DMEM)with 10% calf serum (CS), penicillin and streptomycin. The MSV vectorcontaining the gene is then added to the media and the packaging cellsare transduced with the vector. The packaging cells now produceinfectious viral particles containing the gene (the packaging cells arenow referred to as producer cells).

[0187] Fresh media is added to the transduced producer cells, andsubsequently, the media is harvested from a 10 cm plate of confluentproducer cells. The spent media, containing the infectious viralparticles, is filtered through a millipore filter to remove detachedproducer cells and this media is then used to infect fibroblast cells.Media is removed from a sub-confluent plate of fibroblasts and quicklyreplaced with the media from the producer cells. This media is removedand replaced with fresh media. If the titer of virus is high, thenvirtually all fibroblasts will be infected and no selection is required.If the titer is very low, then it is necessary to use a retroviralvector that has a selectable marker, such as neo or his.

[0188] The engineered fibroblasts are then injected into the host,either alone or after having been grown to confluence on cytodex 3microcarrier beads. The fibroblasts now produce the protein product.Numerous modifications and variations of the present invention arepossible in light of the above teachings and, therefore, within thescope of the appended claims, the invention may be practiced otherwisethan as particularly described.

[0189] This gene is expressed primarily in immune cells, particularlylymphocytes.

[0190] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune orhematopoietic disorders, particularly leukemias. Similarly, polypeptidesand antibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the immune system, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. immune, hematopoietic, and cancerousand wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,urine, synovial fluid or spinal fluid) or another tissue or cell sampletaken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0191] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:69 as residues: Asp-59 to Asn-65, Lys-72 to Trp-79, Tyr-110 toVal-121, Ala-204 to Asn-215.

[0192] The tissue distribution in immune cells, combined with thehomology to a conserved G-protein coupled receptor indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulas a chemokine receptor on lymphocytes that regulate immune response, ormay even be useful in gene therapy to ameliorate the effects of adefective endogenous G-protein, or may show benefit as an antagonist forsuch receptors. The protein can also be used to determine biologicalactivity, to raise antibodies, as tissue markers, to isolate cognateligands or receptors, to identify agents that modulate theirinteractions and as nutritional supplements. It may also have a verywide range of biological acitivities. Typical of these are cytokine,cell proliferation/differentiation modulating activity or induction ofother cytokines; immunostimulating/immunosuppressant activities (e.g.for treating human immunodeficiency virus infection, cancer, autoimmunediseases and allergy); regulation of hematopolesis (e.g. for treatinganaemia or as adjunct to chemotherapy); stimulation or growth of bone,cartilage, tendons, ligaments and/or nerves (e.g. for treating wounds,stimulation of follicle stimulating hormone (for control of fertility);chemotactic and chemokinetic activities (e.g. for treating infections,tumors); hemostatic or thrombolytic activity (e.g. for treatinghaemophelia, cardiac infarction etc.); anti-inflammatory activity (e.g.for treating septic shock, Crohn's disease); as antimicrobials; fortreating psoriasis or other hyperproliferative diseases; for regulationof metabolism, and behaviour. Also contemplated is the use of thecorresponding nucleic acid in gene therapy procedures. Protein, as wellas, antibodies directed against the protein may show utility as a tumormarker and/or immunotherapy targets for the above listed tissues.

[0193] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:33 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 983 of SEQID NO:33, b is an integer of 15 to 997, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:33, and where bis greater than or equal to a+14.

[0194] FEATURES OF PROTEIN ENCODED BY GENE NO: 24

[0195] The translation product of this gene shares sequence homologywith protein disulfide isomerase which is thought to be important inprotein folding and protein-protein interaction. This gene also containsshares homology to genes having thioredoxin domains. (See Accession No.1943817.) This gene also maps to chromosome 9, and therefore may beuseful in linkage analysis as a marker for chromosome 9.

[0196] This gene is expressed primarily in tumor tissues.

[0197] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,proliferative disorders, or disorders involving inappropriate proteinfolding and protein-protein interaction. Similarly, polypeptides andantibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the tumorigenic process, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g., cancerous and wounded tissues) orbodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid orspinal fluid) or another tissue or cell sample taken from an individualhaving such a disorder, relative to the standard gene expression level,i.e., the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0198] Preferred epitopes include those comprising a sequence shown inSEQ ID NO:70 as residues: Glu-78 to Asn-83, Asp-91 to Gln-100, Glu-122to Ser-128, Arg-137 to Pro-143, Asp-157 to Asn-162, Glu-168 to Asn-174,Ser-199 to Gly-206, Pro-213 to Ala-218, Glu-251 to Thr-257, Ser-353 toHis-361, Gly-363 to Ala-375, Pro-382 to Phe-387, Arg-401 to Leu-406.

[0199] The tissue distribution in proliferative tissues, combined withthe homology to a conserved protein disulfide isomerase indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor regulating protein folding and protein-protein interaction in tumortissues. Similarly, expression within cellular sources marked byproliferating cells indicates that this protein may play a role in theregulation of cellular division, and may show utility in the diagnosisand treatment of cancer and other proliferative disorders. Similarly,developmental tissues rely on decisions involving cell differentiationand/or apoptosis in pattern formation. Thus this protein may also beinvolved in apoptosis or tissue differentiation and could again beuseful in cancer therapy. Protein, as well as, antibodies directedagainst the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0200] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:34 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1900 of SEQID NO:34, b is an integer of 15 to 1914, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:34, and whereb is greater than or equal to a+14.

[0201] FEATURES OF PROTEIN ENCODED BY GENE NO: 25

[0202] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: KHTVETRSVAFRKQLNR (SEQ ID NO:189).Polynucleotides encoding these polypeptides are also encompassed by theinvention.

[0203] This gene is expressed primarily in leukocytes involved in immunedefense, including T cells, macrophages, neutrophils, and to a lesserextent, in fetal liver/spleen, synovium, adrenal gland tumor, adipose,and placenta.

[0204] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune orhematopoietic disorders, particularly defects or disorders inleukocytes. Similarly, polypeptides and antibodies directed to thesepolypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theimmune and defense systems, expression of this gene at significantlyhigher or lower levels may be routinely detected in certain tissues orcell types (e.g. immune, hematopoietic, hepatic, reproductive,endocrine, developmental, and cancerous and wounded tissues) or bodilyfluids (e.g., lymph, serum, plasma, urine, synovial fluid or spinalfluid) or another tissue or cell sample taken from an individual havingsuch a disorder, relative to the standard gene expression level, i.e.,the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0205] The tissue distribution in immune cells and tissues indicatesthat polynucleotides and polypeptides corresponding to this gene areuseful for regulating the function of leukocytes, and is useful for thediagnosis and/or treatment of disorders in the immune and defensesystems. This gene product is primarily expressed in hematopoietic cellsand tissues, suggesting that it plays a role in the survival,proliferation, and/or differentiation of hematopoieitic lineages. Thisis particularly supported by the expression of this gene product infetal liver, which is a primary site of definitive hematopoiesis.Expression of this gene product in T cells and macrophage also stronglyindicates a role for this protein in immune function and immunesurveillance. Moreover, This gene product may be involved in theregulation of cytokine production, antigen presentation, or otherprocesses that may also suggest a usefulness in the treatment of cancer(e.g. by boosting immune responses). Since the gene is expressed incells of lymphoid origin, the natural gene product may be involved inimmune functions. Therefore it may be also used as an agent forimmunological disorders including arthritis, asthma, immunodeficiencydiseases such as AIDS, leukemia, rheumatoid arthritis, granulomatousdisease, inflammatory bowel disease, sepsis, acne, neutropenia,neutrophilia, psoriasis, hypersensitivities, such as T-cell mediatedcytotoxicity; immune reactions to transplanted organs and tissues, suchas host-versus-graft and graft-versus-host diseases, or autoimmunitydisorders, such as autoimmune infertility, lense tissue injury,demyelination, systemic lupus erythematosis, drug induced hemolyticanemia, rheumatoid arthritis, Sjogren's disease, scleroderma andtissues. In addition, this gene product may have commercial utility inthe expansion of stem cells and committed progenitors of various bloodlineages, and in the differentiation and/or proliferation of variouscell types. Protein, as well as, antibodies directed against the proteinmay show utility as a tumor marker and/or immunotherapy targets for theabove listed tissues.

[0206] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:35 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1006 of SEQID NO:35, b is an integer of 15 to 1020, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:35, and whereb is greater than or equal to a+14.

[0207] FEATURES OF PROTEIN ENCODED BY GENE NO: 26

[0208] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: PQVLHLRWLPKVLGYRSXPLRLADPSTFXM (SEQID NO:190), QLLGFEGNDSAGERRWRGANMQIPLLQVALPLSTEEGTGPSGPTQPSPQGEVRFLRSPRNIGGQV PHWEWRSHSLPWVLTSTLSGCEGDLPGFPHQVQLPAAESHTLNTGLLRSDTGQFTPCLKLAFERPSG (SEQ ID NO:191), NDSAGERRWRGANMQIPLLQVALP (SEQ ID NO:192), PSPQGEVRFLRSPRMG GQVPHWEWRSHSL(SEQ ID NO:193), and/or HQVQLPAAESHTLNTGLL RSDTGQFTP (SEQ ID NO:194).Polynucleotides encoding these polypeptides are also encompassed by theinvention.

[0209] This gene is expressed in colon cancer, breast cancer,neutrophils, T cells, spinal cord, fibroblasts, and vascular endothelialcells.

[0210] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,gastrointesinal, reproductive, or immune disorders, particularlycancers, and disorder and abnormalities in leukocytes and other tissues.Similarly, polypeptides and antibodies directed to these polypeptidesare useful in providing immunological probes for differentialidentification of the tissue(s) or cell type(s). For a number ofdisorders of the above tissues or cells, particularly those cellsinvolved in tumorigenesis and immune defense systems, expression of thisgene at significantly higher or lower levels may be routinely detectedin certain tissues or cell types (e.g. immune, colon, breast, vascular,and cancerous and wounded tissues) or bodily fluids (e.g., lymph, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0211] The tissue distribution in breast and colon cancer tissuesindicates that polynucleotides and polypeptides corresponding to thisgene are useful for dianosis and treatment of cancer or immune systemdisorders. Furthermore, the tissue distribution in tumors of colon andbreast origins indicates that polynucleotides and polypeptidescorresponding to this gene are useful for diagnosis and intervention ofthese tumors, in addition to other tumors where expression has beenindicated. Furthermore, the protein product of this gene may bebeneficial in the treatment and/or prevention of cancers throughenhancing the immune response to tumor cell surface antigens, or even asa preventative through the correction of an aberrant cell-cycleregulator protein by gene therapy. Protein, as well as, antibodiesdirected against the protein may show utility as a tissue-specificmarker and/or immunotherapy target for the above listed tissues.

[0212] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:36 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 767 of SEQID NO:36, b is an integer of 15 to 781, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:36, and where bis greater than or equal to a+14.

[0213] FEATURES OF PROTEIN ENCODED BY GENE NO: 27

[0214] The translation product of this gene shares sequence homologywith a mouse pancreatic polypeptide (See Accession No. 200464.). Thus,it is likely that this gene has activity similar to the mouse pancreaticpolypeptide. Preferred polypeptide fragments comprise the amino acidssequence: (SEQ ID NO:195)APLETMQNKPRAPQKRALPFPELELRDYASVLTRYSLGLRNKEPSLGHRW GTQKLGRSPC, (SEQ IDNO:196) MQNKPRAPQKRALPFPELELRDYASVLTRYSLGL RNKEPSLGHRWGTQKLGRSPCSEGSQGHTTDAADVQNHSKEEQRDAG AQRXCGQGRHTWAYRXGAQDTSRLTGDPRGGERSPPKCQSMKQQ EGAPSGHCWDQWCHGASEVVWPESRKRAQIFXSPCRQSPRSSALGAGQKL AVCSPDILCCPTDTLLASHPHSLLTGTQFSGQTQALAPSWCA, (SEQ ID NO:197)APQKRALPFPELELRDYASVLTRYSL, (SEQ ID NO:198) APQKRALPFPELELRDYASVLTRYSLG, (SEQ ID NO:199) LGRSPCSEGSQGHTTDAADVQNHS KLEEQR, and/or(SEQ ID NO:200) TDTLLASHPHSLLTGTQFSQTQAL.

[0215] Also preferred are polynucleotide fragments encoding thispolypeptide fragment.

[0216] This gene is expressed primarily in neutrophils, and to a lesserextent, in induced endothelial cells.

[0217] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune,hematopoietic, or vascular disorders, particularly disorders in immunecell adhesion. Similarly, polypeptides and antibodies directed to thesepolypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of theimmune system, expression of this gene at significantly higher or lowerlevels may be routinely detected in certain tissues or cell types (e.g.immune, vascular, and cancerous and wounded tissues) or bodily fluids(e.g. lymph, serum, plasma, urine, synovial fluid or spinal fluid) oranother tissue or cell sample taken from an individual having such adisorder, relative to the standard gene expression level, i.e., theexpression level in healthy tissue or bodily fluid from an individualnot having the disorder.

[0218] The tissue distribution in neutrophils and endothelial cellsindicates that polynucleotides and polypeptides corresponding to thisgene are useful for the regulation of neutrophils or leukocyte adhesionto endothelial cells. It may be used to diagnose or treat disordersassociated with neutrophils and vasular endothelial cells. Furthermore,this gene product may be produced more generally in endothelial cells orwithin the circulation. In such instances, it may play more generalizedroles in vascular function, such as in angiogenesis. It may also beproduced in the vasculature and have effects on other cells within thecirculation, such as hematopoietic cells, or be used as a preventativefor embolisms, atherosclerosis, aneurysms, microvascular disease, orstroke. It may serve to promote the proliferation, survival, activation,and/or differentiation of hematopoietic cells, as well as other cellsthroughout the body. Protein, as well as, antibodies directed againstthe protein may show utility as a tumor marker and/or immunotherapytargets for the above listed tissues.

[0219] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:37 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 952 of SEQID NO:37, b is an integer of 15 to 966, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:37, and where bis greater than or equal to a+14.

[0220] FEATURES OF PROTEIN ENCODED BY GENE NO: 28

[0221] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence:IAQVLKAEMCLVXRPHPXLLDSHRGWAGETLRGQGRQEXESDTKAGTLQLQR QAPLPLTQHSLVLPISPGPSNHTQS (SEQ ID NO:201), RGWAGETLRGQGR QEXESDT (SEQ IDNO:202), and/or APL PLTQHSLVLPISPGPSN (SEQ ID NO:203). Polynucleotidesencoding these polypeptides are also encompassed by the invention.

[0222] This gene is expressed primarily in prostate BPH.

[0223] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,proliferative or reproductive disorders, particularly benign hypertrophyof the prostate. Similarly, polypeptides and antibodies directed tothese polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of themale urogenital system, expression of this gene at significantly higheror lower levels may be routinely detected in certain tissues or celltypes (e.g. prostate, urogenital, and cancerous and wounded tissues) orbodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid orspinal fluid) or another tissue or cell sample taken from an individualhaving such a disorder, relative to the standard gene expression level,i.e., the expression level in healthy tissue or bodily fluid from anindividual not having the disorder.

[0224] The tissue distribution in prostate tissue indicates thatpolynucleotides and polypeptides corresponding to this gene are usefulfor the diagnosis and treatment of benign or metastatic hypertrophy ofthe prostate or prostate cancer. Expression within cellular sourcesmarked by proliferating cells indicates that this protein may play arole in the regulation of cellular division, and may show utility in thediagnosis and treatment of cancer and other proliferative disorders.Similarly, developmental tissues rely on decisions involving celldifferentiation and/or apoptosis in pattern formation. Thus this proteinmay also be involved in apoptosis or tissue differentiation and couldagain be useful in cancer therapy. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0225] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID INO:38 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 402 of SEQID NO:38, b is an integer of 15 to 416, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:38, and where bis greater than or equal to a+14.

[0226] FEATURES OF PROTEIN ENCODED BY GENE NO: 29

[0227] The translation product of this gene shares sequence homologywith C16C10.7, a C. elegans gene similar to zinc finger protein, aprotein involved in DNA binding. Thus, this protein is expected to sharecertain biological activities with C1610.7 including DNA bindingactivities. One embodiment of this gene comprises polypeptides of thefollowing amino acid sequence: (SEQ ID NO:204)NRERGGAGATFECNICLETAREAVVSVCGHLYCWPCLHQWLETRPERQECPVCKAGISREKVVPLYGRGSQKQDPRLKTPPRPQGQRPAPESRGGFQPFGDTGGFHFSFGVGAFPFGFFTTVFNAHEPFRRGTGVDLGQGHPASSWQDSL FLFLAIFFFFWLLSI, (SEQID NO:205) NRERGGAGATFECNICLETAREAVVSVCGHLYCWPCLHQWLETRPERQECPVCKAGISREKVVPLYGRGSQKPQDPRLKTPPRPQGQRPAPESRGGFQPFGDTGGFHFSFGVGAFPFGFFTTVFNAHEPFRRGTG VDLGQGHPASSWQD, (SEQID NO:206) NRERGGAGATFECNICLETAREAVVSVC GHLYCWPCLHQWL, (SEQ ID NO:207)ETRPERQECPVCKAGISREKVVPLYG RGSQKPQDPRLK, (SEQ ID NO:208)TPPRPQGQRPAPESRGGFQPFGDTGGFH FSFGVG, (SEQ ID NO:209)AFPFGFFTTVFNAHEPFRRGTGVDLGQGHPAS SWQD.

[0228] An additional embodiment is the polynucleotides encoding thesepolypeptides.

[0229] This gene is expressed primarily in activated T-cells, and to alesser extent, in fetal brain, TNF-induced amniotic cells andepididymus.

[0230] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to, immune,neurodegenerative, or developmental disorders. Similarly, polypeptidesand antibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the immune and central nervous systems, expression ofthis gene at significantly higher or lower levels may be routinelydetected in certain tissues or cell types (e.g. immune, neural,developmental, reproductive, and cancerous and wounded tissues) orbodily fluids (e.g., lymph, amniotic fluid, seminal fluid, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0231] The tissue distribution in T-cells and fetal brain indicates thatthe protein products of this gene are useful for the diagnosis andtreatment of immune and/or neurodegenerative disorders, as well asuseful for the promotion of, survival, and differentiation of neurons.Furthermore, the tissue distribution indicates that polynucleotides andpolypeptides corresponding to this gene are useful for the diagnosisand/or treatment of hematopoietic disorders. Expression of this geneproduct in T cells also strongly indicates a role for this protein inimmune function and immune surveillance. Expression within embryonictissue and other cellular sources marked by proliferating cellsindicates that this protein may play a role in the regulation ofcellular division, and may show utility in the diagnosis and treatmentof cancer and other proliferative disorders. Similarly, developmentaltissues rely on decisions involving cell differentiation and/orapoptosis in pattern formation. Thus this protein may also be involvedin apoptosis or tissue differentiation and could again be useful incancer therapy. Protein, as well as, antibodies directed against theprotein may show utility as a tumor marker and/or immunotherapy targetsfor the above listed tissues.

[0232] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:39 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 1100 of SEQID NO:39, b is an integer of 15 to 1114, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:39, and whereb is greater than or equal to a+14.

[0233] FEATURES OF PROTEIN ENCODED BY GENE NO: 30

[0234] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: GLSTGPDMASLDLFV (SEQ ID NO:210),GRPTRPSQAT RHFLLGTLFTNCLCGTFCFPCLGCQVAADMNECCLCGTSVAMRTLYRTRYGIP GSICDDYMATLCCPHCTLCQIKRDINRRRAMRTF (SEQ ID NO:211), IKNLIFFMPSVVLKHIHHISVAKDGEELKLKRCLLNFVASVRAFHHQFLESTHGSPSVDISLDLAKSTMRTAKSCHIVITNRSRDA ISGPVESPHCDACSTQTAFIHISCNLTPKARETKCATETISKQGSEQEMSCGLGRTRGST (SEQ ID NO:212), FLLGTLFTNCLCGTFCFPCLGCQ (SEQ ID NO:213), SICDDY MATLCCPHCTLCQIKRDI (SEQ IDNO:214), SVVLKHIHHISVAKDGEELKLKRCLLNFVA (SEQ ID NO:215),NFVASVRAFHHQFLESTHGSPSVDIS (SEQ ID NO:216), and/or TAFIHISCNLTPKARETKCATETISKQG (SEQ ID NO:217). Polynucleotides encoding thesepolypeptides are also encompassed by the invention. The gene encodingthe disclosed cDNA is thought to reside on chromosome 4. Accordingly,polynucleotides related to this invention are useful as a marker inlinkage analysis for chromosome 4.

[0235] This gene is expressed primarily in T-cells, fetal tissues, andplacenta, and to a lesser extent in bone marrow.

[0236] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,immunological, developmental, or reproductive disorders, includingautoimmune diseases or congenital defects. Similarly, polypeptides andantibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the immune system, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. immune, develpomental, placental,hematopoietic, and cancerous and wounded tissues) or bodily fluids (e.g.lymph, serum, plasma, urine, synovial fluid or spinal fluid) or anothertissue or cell sample taken from an individual having such a disorder,relative to the standard gene expression level, i.e., the expressionlevel in healthy tissue or bodily fluid from an individual not havingthe disorder.

[0237] The tissue distribution in T-cells and bone marrow indicates thatthe protein products of this gene are useful for diagnosis and treatmentof immunologically mediated disorders as they are thought to play a rolein the proliferation, survival, differentiation, and/or activation of avariety of hematopoietic cells, including early progenitors orhematopoietic stem cells. Furthermore, the tissue distribution indicatesthat polynucleotides and polypeptides corresponding to this gene areuseful for the diagnosis and treatment of cancer and other proliferativedisorders. Expression within embryonic tissue and other cellular sourcesmarked by proliferating cells indicates that this protein may play arole in the regulation of cellular division. Additionally, theexpression in hematopoietic cells and tissues indicates that thisprotein may play a role in the proliferation, differentiation, and/orsurvival of hematopoietic cell lineages. In such an event, this gene maybe useful in the treatment of lymphoproliferative disorders, and in themaintenance and differentiation of various hematopoietic lineages fromearly hematopoietic stem and committed progenitor cells. Similarly,embryonic development also involves decisions involving celldifferentiation and/or apoptosis in pattern formation. Thus, thisprotein may also be involved in apoptosis or tissue differentiation andcould again be useful in cancer therapy. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker and/orimmunotherapy targets for the above listed tissues.

[0238] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:40 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 588 of SEQID NO:40, b is an integer of 15 to 602, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:40, and where bis greater than or equal to a+14.

[0239] FEATURES OF PROTEIN ENCODED BY GENE NO: 31

[0240] In specific embodiments, polypeptides of the invention comprisethe following amino acid sequence: MKGEIE (SEQ ID NO:218).Polynucleotides encoding these polypeptides are also encompassed by theinvention.

[0241] This gene is expressed primarily in human skin, and to a lesserextent, in fetal heart tissue.

[0242] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,integumentary, developmental, or cardiovascular disorders, particularlywound healing conditions and skin cancers. Similarly, polypeptides andantibodies directed to these polypeptides are useful in providingimmunological probes for differential identification of the tissue(s) orcell type(s). For a number of disorders of the above tissues or cells,particularly of the integumentary system, expression of this gene atsignificantly higher or lower levels may be routinely detected incertain tissues or cell types (e.g. skin, developmental, cardiovascular,and cancerous and wounded tissues) or bodily fluids (e.g., lymph, serum,plasma, urine, synovial fluid or spinal fluid) or another tissue or cellsample taken from an individual having such a disorder, relative to thestandard gene expression level, i.e., the expression level in healthytissue or bodily fluid from an individual not having the disorder.

[0243] The tissue distribution in skin indicates that the proteinproducts of this gene are useful for diagnosis and treatment of skincancers and wound healing. Moreover, polynucleotides and polypeptidescorresponding to this gene are useful for the treatment, diagnosis,and/or prevention of various skin disorders including congenitaldisorders (i.e. nevi, moles, freckles, Mongolian spots, hemangiomas,port-wine syndrome), integumentary tumors (i.e. keratoses, Bowen'sdisease, basal cell carcinoma, squamous cell carcinoma, malignantmelanoma, Paget's disease, mycosis fungnoides, and Kaposi's sarcoma),injuries and inflammation of the skin (i.e.wounds, rashes, prickly heatdisorder, psoriasis, dermatitis), atherosclerosis, uticaria, eczema,photosensitivity, autoimmune disorders (i.e. lupus erythematosus,vitiligo, dermnatomyositis, morphea, scleroderma, pemphigoid, andpemphigus), keloids, striae, erythema, petechiae, purpura, andxanthelasma. In addition, such disorders may predispose increasedsusceptibility to viral and bacterial infections of the skin (i.e. coldsores, warts, chickenpox, molluscum contagiosum, herpes zoster, boils,cellulitis, erysipelas, impetigo, tinea, althletes foot, and ringworm).

[0244] Moreover, the protein product of this gene may also be useful forthe treatment or diagnosis of various connective tissue disorders suchas arthritis, trauma, tendonitis, chrondomalacia and inflammation,autoimmune disorders such as rheumatoid arthritis, lupus, scleroderma,and dermatomyositis as well as dwarfism, spinal deformation, andspecific joint abnormalities as well as chondrodysplasias (i.e.spondyloepiphyseal dysplasia congenita, familial osteoarthritis,Atelosteogenesis type II, metaphyseal chondrodysplasia type Schmid).Additionally, it may play more generalized roles in vascular function,such as in angiogenesis. It may also be produced in the vasculature andhave effects on other cells within the circulation, such ashematopoietic cells. It may serve to promote the proliferation,survival, activation, and/or differentiation of hematopoietic cells, aswell as other cells throughout the body. Protein, as well as, antibodiesdirected against the protein may show utility as a tumor marker andlorimmunotherapy targets for the above listed tissues.

[0245] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:41 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 956 of SEQID NO:41, b is an integer of 15 to 970, where both a and b correspond tothe positions of nucleotide residues shown in SEQ ID NO:41, and where bis greater than or equal to a+14.

[0246] FEATURES OF PROTEIN ENCODED BY GENE NO: 32

[0247] The translation product of this gene shares sequence homologywith human Tear Prealbumin (GenBank accession no. gil1307518) and ratOderant-binding protein (GenBank accession no. gil90755 1), both ofwhich are thought to be important in molecule binding and transport. Thetranslation product of this gene is a homolog of the lipocalin family,which are thought to be involved in the transport of small hydrophobicmolecules. In specific embodiments, polypeptides of the inventioncomprise the following amino acid sequence: EFGTSRGRQHRALE (SEQ IDNO:219), HQTP GVTGLSAVEMDQITPALWEALAIDTLRKLRIGTRRPRIRWGQEAHV PAGAAQEGPLHLLLQRPAPWGXAPHGKACG (SEQ ID NO:220), GLGQGGQGLDGGRK LMYLQELPRRDHYIFYCKDQHHGGXLHMGKLVGRNSDTNREALEEFKK LVQRKGLSEEDIFTPLQTGSCVPEH(SEQ ID NO:221), SGPSRLRTSLSHPVS DVRATSPPGRRGQPLLGGGQSWGPGKRAAWALSTCGGWCTGVGGGG XWGWEWGRGSQALYLPGSSVFRXRIFFWMHRSSLMKVNVASNFPPPRAVTF TGDTFWASCLR KYLSTTMAFTYQVPVISSSXRVKDRAAAXPSVTPRNRV FISRALCCRPRLVPN (SEQ IDNO:222), GLPEGRRDLVHLDCGQACHTRCLMS GPPAPQEGEAS PSLEVGRAGALAKGQPGHSLPVEAGALGLAVGEGGGGXG GGAHRRCICQAPPSSAXGFSSGCTDPPS (SEQ ID NO:223), VEMDQITPALWEALAIDTLRKLRIGTRRPR (SEQ ID NO:224), RKLMY LQELPRRDHYIFYCK DQH (SEQ IDNO:225), EALEEFKKLVQRKGLSEEDIFTP (SEQ ID NO:226), RATSPPGRRGQPLLGGGQSWGPGKRAA (SEQ ID NO:227), FFWMHRSSLMKV NVASNFPPPRAVTFTGD(SEQ ID NO:228), and/or CLMSGPPAPQEGEASP SLEVGRAGALAK (SEQ ID NO:229).Polynucleotides encoding these polypeptides are also encompassed by theinvention.

[0248] This gene is expressed primarily in endometrial tumor.

[0249] Therefore, polynucleotides and polypeptides of the invention areuseful as reagents for differential identification of the tissue(s) orcell type(s) present in a biological sample and for diagnosis ofdiseases and conditions which include, but are not limited to,reproductive disorders, particularly cancers of the endometrium, skinand haemopoietic system. Similarly, polypeptides and antibodies directedto these polypeptides are useful in providing immunological probes fordifferential identification of the tissue(s) or cell type(s). For anumber of disorders of the above tissues or cells, particularly of thehaemopoietic system, expression of this gene at significantly higher orlower levels may be routinely detected in certain tissues or cell types(e.g. immune, reproductive, and cancerous and wounded tissues) or bodilyfluids (e.g., lymph, serum, amniotic fluid, plasma, urine, synovialfluid or spinal fluid) or another tissue or cell sample taken from anindividual having such a disorder, relative to the standard geneexpression level, i.e., the expression level in healthy tissue or bodilyfluid from an individual not having the disorder.

[0250] The tissue distribution in endometrium tissue, combined with thehomology to the molecule binding and transport gene family indicatesthat the protein products of this gene are useful for the diagnosis andtreatment of cancers of the endometrium and haemopoietic system as wellas for the treatment of autoimmune disorders such as inflammation.Protein, as well as, antibodies directed against the protein may showutility as a tumor marker and/or immunotherapy targets for the abovelisted tissues.

[0251] Many polynucleotide sequences, such as EST sequences, arepublicly available and accessible through sequence databases. Some ofthese sequences are related to SEQ ID NO:42 and may have been publiclyavailable prior to conception of the present invention. Preferably, suchrelated polynucleotides are specifically excluded from the scope of thepresent invention. To list every related sequence is cumbersome.Accordingly, preferably excluded from the present invention are one ormore polynucleotides comprising a nucleotide sequence described by thegeneral formula of a-b, where a is any integer between 1 to 988 of SEQID NO:42, b is an integer of 15 to 1002, where both a and b correspondto the positions of nucleotide residues shown in SEQ ID NO:42, and whereb is greater than or equal to a+14. 5′ NT of AA First Last ATCC NT 5′ NT3′ NT 5′ NT First SEQ AA AA First Last Deposit SEQ Total of of of AA ofID of of AA of AA cDNA Nr ID NT Clone Clone Start Signal NO: Sig SigSecreted of Gene No. Clone ID and Date Vector NO: X Seq. Seq. Seq. CodonPep Y Pep Pep Portion ORF 1 HSVBZ80  97978 Uni-ZAP XR 11 1169 64 1060162 162 47 1 38 39 145 Mar. 27, 1997 209075 May 22, 1997 2 HTAAU21 97978 Uni-ZAP XR 12 1310 1 1310 283 283 48 1 18 19 311 Mar. 27, 1997209075 May 22, 1997 3 HTLEK16  97978 Uni-ZAP XR 13 1139 19 1111 251 49 121 22 46 Mar. 27, 1997 209075 May 22, 1997 4 HUSIR91  97978 pSport1 142271 743 2271 59 59 50 1 23 24 467 Mar. 27, 1997 209075 May 22, 1997 4HUSIR91  97978 pSport1 43 2581 1035 2164 1148 1148 79 1 27 28 207 Mar.27, 1997 209075 May 22, 1997 5 HADMC21  97978 pBluescript 15 626 60 47991 91 51 1 51 52 82 Mar. 27, 1997 209075 May 22, 1997 6 HAGFM45  97978Uni-ZAP XR 16 2118 1170 2058 1248 1248 52 1 16 17 62 Mar. 27, 1997209075 May 22, 1997 7 HAIBE65  97978 Uni-ZAP XR 17 1076 396 993 528 52853 1 31 32 123 Mar. 27, 1997 209075 May 22, 1997 8 HAQBH57  97978Uni-ZAP XR 18 1379 420 1306 618 618 54 1 25 26 179 Mar. 27, 1997 209075May 22, 1997 9 HATCX80  97978 Uni-ZAP XR 19 1337 47 1337 199 199 55 1 1819 286 Mar. 27, 1997 209075 May 22, 1997 10 HCFLQ84  97978 pSport1 201390 237 1390 410 410 56 1 20 21 33 Mar. 27, 1997 209075 May 22, 1997 11HCFLS78  97978 pSport1 21 1431 178 981 420 420 57 1 21 22 23 Mar. 27,1997 209075 May 22, 1997 12 HTADI12  97978 Uni-ZAP XR 22 2539 69 2539104 104 58 1 27 28 46 Mar. 27, 1997 209075 May 22, 1997 13 HEMCM42 97978 Uni-ZAP XR 23 1041 48 1007 58 58 59 1 29 30 113 Mar. 27, 1997209075 May 22, 1997 14 HEONP72  97978 pSport1 24 1962 1 1947 181 181 601 19 20 31 Mar. 27, 1997 209075 May 22, 1997 15 HFCDW34  97978 Uni-ZAPXR 25 1228 321 1228 525 525 61 1 24 25 80 Mar. 27, 1997 209075 May 22,1997 16 HTTEU91  97978 Uni-ZAP XR 26 1340 325 1340 15 15 62 1 18 19 104Mar. 27, 1997 209075 May 22, 1997 17 HHGBF89  97978 Lambda ZAP 27 806 31806 77 77 63 1 19 20 145 Mar. 27, 1997 II 209075 May 22, 1997 17 HHGBF89 97978 Lambda ZAP 44 796 31 796 77 77 80 1 25 26 145 Mar. 27, 1997 II209075 May 22, 1997 18 HKIYQ65  97978 pBluescript 28 696 1 684 98 98 641 17 18 30 Mar. 27, 1997 209075 May 22, 1997 19 HKMLN27  97978pBluescript 29 1007 71 963 129 129 65 1 23 24 259 Mar. 27, 1997 209075May 22, 1997 20 HKIAC30 209022 Uni-ZAP XR 30 2026 131 2018 166 166 66 130 31 339 May 08, 1997 20 HKIAC30 209022 Uni-ZAP XR 45 2017 126 2007 161161 81 1 22 May 08, 1997 21 HKIXB95 209022 pBluescript 31 699 196 699230 230 67 1 22 23 26 May 08, 1997 22 HLMIY86 209022 Lambda ZAP 32 12641 1264 560 560 68 1 24 25 76 May 08, 1997 II 23 HLYAZ61 209022 pSport133 997 74 997 205 205 69 1 20 21 215 May 08, 1997 24 HMQDT36 209022Uni-ZAP XR 34 1914 37 1897 192 192 70 1 32 33 406 May 08, 1997 25HNEDF25 209022 Uni-ZAP XR 35 1020 1 1010 210 210 71 1 13 14 45 May 08,1997 26 HNFET17 209022 Uni-ZAP XR 36 781 31 781 100 100 72 1 33 May 08,1997 27 HNHCR46 209022 Uni-ZAP XR 37 966 19 460 458 458 73 1 24 25 160May 08, 1997 28 HPWAS91 209022 Uni-ZAP XR 38 416 1 416 95 95 74 1 24 2525 May 08, 1997 29 HWTAW41 209022 Uni-ZAP XR 39 1114 804 1114 50 50 75 143 44 91 May 08, 1997 30 HBMUT52 209022 Uni-ZAP XR 40 602 142 602 204204 76 1 26 27 32 May 08, 1997 31 HERAG83 209022 Uni-ZAP XR 41 970 1 970110 110 77 1 22 23 22 May 08, 1997 32 HETFI51 209022 Uni-ZAP XR 42 10021 1002 43 43 78 1 21 22 173 May 08, 1997 32 HETFI51 209022 Uni-ZAP XR 46981 1 981 23 23 82 1 17 18 30 May 08, 1997

[0252] Table 1 summarizes the information corresponding to each “GeneNo.” described above. The nucleotide sequence identified as “NT SEQ IDNO:X” was assembled from partially homologous (“overlapping”) sequencesobtained from the “cDNA clone ID” identified in Table 1 and, in somecases, from additional related DNA clones. The overlapping sequenceswere assembled into a single contiguous sequence of high redundancy(usually three to five overlapping sequences at each nucleotideposition), resulting in a final sequence identified as SEQ ID NO:X.

[0253] The cDNA Clone ID was deposited on the date and given thecorresponding deposit number listed in “ATCC Deposit No:Z and Date.”Some of the deposits contain multiple different clones corresponding tothe same gene. “Vector” refers to the type of vector contained in thecDNA Clone ID.

[0254] “Total NT Seq.” refers to the total number of nucleotides in thecontig identified by “Gene No.” The deposited clone may contain all ormost of these sequences, reflected by the nucleotide position indicatedas “5′ NT of Clone Seq.” and the “3′ NT of Clone Seq.” of SEQ ID NO:X.The nucleotide position of SEQ ID NO:X of the putative start codon(methionine) is identified as “5′ NT of Start Codon.” Similarly, thenucleotide position of SEQ ID NO:X of the predicted signal sequence isidentified as “5′ NT of First AA of Signal Pep.”

[0255] The translated amino acid sequence, beginning with themethionine, is identified as “AA SEQ ID NO:Y,” although other readingframes can also be easily translated using known molecular biologytechniques. The polypeptides produced by these alternative open readingframes are specifically contemplated by the present invention.

[0256] The first and last amino acid position of SEQ ID NO:Y of thepredicted signal peptide is identified as “First AA of Sig Pep” and“Last AA of Sig Pep.” The predicted first amino acid position of SEQ IDNO:Y of the secreted portion is identified as “Predicted First AA ofSecreted Portion.” Finally, the amino acid position of SEQ ID NO:Y ofthe last amino acid in the open reading frame is identified as “Last AAof ORF.”

[0257] SEQ ID NO:X and the translated SEQ ID NO:Y are sufficientlyaccurate and otherwise suitable for a variety of uses well known in theart and described further below. For instance, SEQ ID NO:X is useful fordesigning nucleic acid hybridization probes that will detect nucleicacid sequences contained in SEQ ID NO:X or the cDNA contained in thedeposited clone. These probes will also hybridize to nucleic acidmolecules in biological samples, thereby enabling a variety of forensicand diagnostic methods of the invention. Similarly, polypeptidesidentified from SEQ ID NO:Y may be used to generate antibodies whichbind specifically to the secreted proteins encoded by the cDNA clonesidentified in Table 1.

[0258] Nevertheless, DNA sequences generated by sequencing reactions cancontain sequencing errors. The errors exist as misidentifiednucleotides, or as insertions or deletions of nucleotides in thegenerated DNA sequence. The erroneously inserted or deleted nucleotidescause frame shifts in the reading frames of the predicted amino acidsequence. In these cases, the predicted amino acid sequence divergesfrom the actual amino acid sequence, even though the generated DNAsequence may be greater than 99.9% identical to the actual DNA sequence(for example, one base insertion or deletion in an open reading frame ofover 1000 bases).

[0259] Accordingly, for those applications requiring precision in thenucleotide sequence or the amino acid sequence, the present inventionprovides not only the generated nucleotide sequence identified as SEQ IDNO:X and the predicted translated amino acid sequence identified as SEQID NO:Y, but also a sample of plasmid DNA containing a human cDNA of theinvention deposited with the ATCC, as set forth in Table 1. Thenucleotide sequence of each deposited clone can readily be determined bysequencing the deposited clone in accordance with known methods. Thepredicted amino acid sequence can then be verified from such deposits.Moreover, the amino acid sequence of the protein encoded by a particularclone can also be directly determined by peptide sequencing or byexpressing the protein in a suitable host cell containing the depositedhuman cDNA, collecting the protein, and determining its sequence.

[0260] The present invention also relates to the genes corresponding toSEQ ID NO:X, SEQ ID NO:Y, or the deposited clone. The corresponding genecan be isolated in accordance with known methods using the sequenceinformation disclosed herein. Such methods include preparing probes orprimers from the disclosed sequence and identifying or amplifying thecorresponding gene from appropriate sources of genomic material.

[0261] Also provided in the present invention are species homologs.Species homologs may be isolated and identified by making suitableprobes or primers from the sequences provided herein and screening asuitable nucleic acid source for the desired homologue.

[0262] The polypeptides of the invention can be prepared in any suitablemanner. Such polypeptides include isolated naturally occurringpolypeptides, recombinantly produced polypeptides, syntheticallyproduced polypeptides, or polypeptides produced by a combination ofthese methods. Means for preparing such polypeptides are well understoodin the art.

[0263] The polypeptides may be in the form of the secreted protein,including the mature form, or may be a part of a larger protein, such asa fusion protein (see below). It is often advantageous to include anadditional amino acid sequence which contains secretory or leadersequences, pro-sequences, sequences which aid in purification, such asmultiple histidine residues, or an additional sequence for stabilityduring recombinant production.

[0264] The polypeptides of the present invention are preferably providedin an isolated form, and preferably are substantially purified. Arecombinantly produced version of a polypeptide, including the secretedpolypeptide, can be substantially purified by the one-step methoddescribed in Smith and Johnson, Gene 67:31-40 (1988). Polypeptides ofthe invention also can be purified from natural or recombinant sourcesusing antibodies of the invention raised against the secreted protein inmethods which are well known in the art.

[0265] Signal Sequences

[0266] Methods for predicting whether a protein has a signal sequence,as well as the cleavage point for that sequence, are available. Forinstance, the method of McGeoch, Virus Res. 3:271-286 (1985), uses theinformation from a short N-termninal charged region and a subsequentuncharged region of the complete (uncleaved) protein. The method of vonHeinje, Nucleic Acids Res. 14:4683-4690 (1986) uses the information fromthe residues surrounding the cleavage site, typically residues −13 to+2, where +1 indicates the amino terminus of the secreted protein. Theaccuracy of predicting the cleavage points of known mammalian secretoryproteins for each of these methods is in the range of 75-80%. (vonHeinje, supra.) However, the two methods do not always produce the samepredicted cleavage point(s) for a given protein.

[0267] In the present case, the deduced amino acid sequence of thesecreted polypeptide was analyzed by a computer program called SignalP(Henrik Nielsen et al., Protein Engineering 10:1-6 (1997)), whichpredicts the cellular location of a protein based on the amino acidsequence. As part of this computational prediction of localization, themethods of McGeoch and von Heinje are incorporated. The analysis of theamino acid sequences of the secreted proteins described herein by thisprogram provided the results shown in Table 1.

[0268] As one of ordinary skill would appreciate, however, cleavagesites sometimes vary from organism to organism and cannot be predictedwith absolute certainty. Accordingly, the present invention providessecreted polypeptides having a sequence shown in SEQ ID NO:Y which havean N-terminus beginning within 5 residues (i.e., +or −5 residues) of thepredicted cleavage point. Similarly, it is also recognized that in somecases, cleavage of the signal sequence from a secreted protein is notentirely uniform, resulting in more than one secreted species. Thesepolypeptides, and the polynucleotides encoding such polypeptides, arecontemplated by the present invention.

[0269] Moreover, the signal sequence identified by the above analysismay not necessarily predict the naturally occurring signal sequence. Forexample, the naturally occurring signal sequence may be further upstreamfrom the predicted signal sequence. However, it is likely that thepredicted signal sequence will be capable of directing the secretedprotein to the ER. These polypeptides, and the polynucleotides encodingsuch polypeptides, are contemplated by the present invention.

[0270] Polynucleotide and Polypeptide Variants

[0271] “Variant” refers to a polynucleotide or polypeptide differingfrom the polynucleotide or polypeptide of the present invention, butretaining essential properties thereof. Generally, variants are overallclosely similar, and, in many regions, identical to the polynucleotideor polypeptide of the present invention.

[0272] By a polynucleotide having a nucleotide sequence at least, forexample, 95% “identical” to a reference nucleotide sequence of thepresent invention, it is intended that the nucleotide sequence of thepolynucleotide is identical to the reference sequence except that thepolynucleotide sequence may include up to five point mutations per each100 nucleotides of the reference nucleotide sequence encoding thepolypeptide. In other words, to obtain a polynucleotide having anucleotide sequence at least 95% identical to a reference nucleotidesequence, up to 5% of the nucleotides in the reference sequence may bedeleted or substituted with another nucleotide, or a number ofnucleotides up to 5% of the total nucleotides in the reference sequencemay be inserted into the reference sequence. The query sequence may bean entire sequence shown in Table 1, the ORF (open reading frame), orany fragement specified as described herein.

[0273] As a practical matter, whether any particular nucleic acidmolecule or polypeptide is at least 90%, 95%, 96%, 97%, 98% or 99%identical to a nucleotide sequence of the presence invention can bedetermined conventionally using known computer programs. A preferredmethod for determing the best overall match between a query sequence (asequence of the present invention) and a subject sequence, also referredto as a global sequence alignment, can be determined using the FASTDBcomputer program based on the algorithm of Brutlag et al. (Comp. App.Biosci. (1990) 6:237-245). In a sequence alignment the query and subjectsequences are both DNA sequences. An RNA sequence can be compared byconverting U's to T's. The result of said global sequence alignment isin percent identity. Preferred parameters used in a FASTDB alignment ofDNA sequences to calculate percent identiy are: Matrix=Unitary,k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization GroupLength=0, Cutoff Score=5, Gap Penalty=5, Gap Size Penalty 0.05, WindowSize=500 or the lenght of the subject nucleotide sequence, whichever isshorter.

[0274] If the subject sequence is shorter than the query sequencebecause of 5′ or 3′ deletions, not because of internal deletions, amanual correction must be made to the results. This is becuase theFASTDB program does not account for 5′ and 3′ truncations of the subjectsequence when calculating percent identity. For subject sequencestruncated at the 5′ or 3′ ends, relative to the the query sequence, thepercent identity is corrected by calculating the number of bases of thequery sequence that are 5′ and 3′ of the subject sequence, which are notmatched/aligned, as a percent of the total bases of the query sequence.Whether a nucleotide is matched/aligned is determined by results of theFASTDB sequence alignment. This percentage is then subtracted from thepercent identity, calculated by the above FASTDB program using thespecified parameters, to arrive at a final percent identity score. Thiscorrected score is what is used for the purposes of the presentinvention. Only bases outside the 5′ and 3′ bases of the subjectsequence, as displayed by the FASTDB alignment, which are notmatched/aligned with the query sequence, are calculated for the purposesof manually adjusting the percent identity score.

[0275] For example, a 90 base subject sequence is aligned to a 100 basequery sequence to determine percent identity. The deletions occur at the5′ end of the subject sequence and therefore, the FASTDB alignment doesnot show a matched/alignement of the first 10 bases at 5′ end. The 10unpaired bases represent 10% of the sequence (number of bases at the 5′and 3′ ends not matched/total number of bases in the query sequence) so10% is subtracted from the percent identity score calculated by theFASTDB program. If the remaining 90 bases were perfectly matched thefinal percent identity would be 90%. In another example, a 90 basesubject sequence is compared with a 100 base query sequence. This timethe deletions are internal deletions so that there are no bases on the5′ or 3′ of the subject sequence which are not matched/aligned with thequery. In this case the percent identity calculated by FASTDB is notmanually corrected. Once again, only bases 5′ and 3′ of the subjectsequence which are not matched/aligned with the query sequence aremanually corrected for. No other manual corrections are to made for thepurposes of the present invention.

[0276] By a polypeptide having an amino acid sequence at least, forexample, 95% “identical” to a query amino acid sequence of the presentinvention, it is intended that the amino acid sequence of the subjectpolypeptide is identical to the query sequence except that the subjectpolypeptide sequence may include up to five amino acid alterations pereach 100 amino acids of the query amino acid sequence. In other words,to obtain a polypeptide having an amino acid sequence at least 95%identical to a query amino acid sequence, up to 5% of the amino acidresidues in the subject sequence may be inserted, deleted, (indels) orsubstituted with another amino acid. These alterations of the referencesequence may occur at the amino or carboxy terminal positions of thereference amino acid sequence or anywhere between those terminalpositions, interspersed either individually among residues in thereference sequence or in one or more contiguous groups within thereference sequence.

[0277] As a practical matter, whether any particular polypeptide is atleast 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, theamino acid sequences shown in Table 1 or to the amino acid sequenceencoded by deposited DNA clone can be determined conventionally usingknown computer programs. A preferred method for determing the bestoverall match between a query sequence (a sequence of the presentinvention) and a subject sequence, also referred to as a global sequencealignment, can be determined using the FASTDB computer program based onthe algorithm of Brutlag et al. (Comp. App. Biosci. (1990) 6:237-245).In a sequence alignment the query and subject sequences are either bothnucleotide sequences or both amino acid sequences. The result of saidglobal sequence alignment is in percent identity. Preferred parametersused in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2,Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0,Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap SizePenalty=0.05, Window Size=500 or the length of the subject amino acidsequence, whichever is shorter.

[0278] If the subject sequence is shorter than the query sequence due toN- or C-terminal deletions, not because of internal deletions, a manualcorrection must be made to the results. This is becuase the FASTDBprogram does not account for N- and C-terminal truncations of thesubject sequence when calculating global percent identity. For subjectsequences truncated at the N- and C-terminal, relative to the the querysequence, the percent identity is corrected by calculating the number ofresidues of the query sequence that are N- and C-terminal of the subjectsequence, which are not matched/aligned with a corresponding subjectresidue, as a percent of the total bases of the query sequence. Whethera residue is matched/aligned is determined by results of the FASTDBsequence alignment. This percentage is then subtracted from the percentidentity, calculated by the above FASTDB program using the specifiedparameters, to arrive at a final percent identity score. This finalpercent identity score is what is used for the purposes of the presentinvention. Only residues to the N- and C-termini of the subjectsequence, which are not matched/aligned with the query sequence, areconsidered for the purposes of manually adjusting the percent identityscore. That is, only query residue positions outside the farthest N- andC-terminal residues of the subject sequence.

[0279] For example, a 90 amino acid residue subject sequence is alignedwith a 100 residue query sequence to determine percent identity. Thedeletion occurs at the N-terminus of the subject sequence and therefore,the FASTDB alignment does not show a matching/alignment of the first 10residues at the N-terminus. The 10 unpaired residues represent 10% ofthe sequence (number of residues at the N- and C-termini notmatched/total number of residues in the query sequence) so 10% issubtracted from the percent identity score calculated by the FASTDBprogram. If the remaining 90 residues were perfectly matched the finalpercent identity would be 90%. In another example, a 90 residue subjectsequence is compared with a 100 residue query sequence. This time thedeletions are internal deletions so there are no residues at the N- orC-termini of the subject sequence which are not matched/aligned with thequery. In this case the percent identity calculated by FASTDB is notmanually corrected. Once again, only residue positions outside the N-and C-terminal ends of the subject sequence, as displayed in the FASTDBalignment, which are not matched/aligned with the query sequnce aremanually corrected for. No other manual corrections are to made for thepurposes of the present invention.

[0280] The variants may contain alterations in the coding regions,non-coding regions, or both. Especially preferred are polynucleotidevariants containing alterations which produce silent substitutions,additions, or deletions, but do not alter the properties or activitiesof the encoded polypeptide. Nucleotide variants produced by silentsubstitutions due to the degeneracy of the genetic code are preferred.Moreover, variants in which 5-10, 1-5, or 1-2 amino acids aresubstituted, deleted, or added in any combination are also preferred.Polynucleotide variants can be produced for a variety of reasons, e.g.,to optimize codon expression for a particular host (change codons in thehuman mRNA to those preferred by a bacterial host such as E. coli).

[0281] Naturally occurring variants are called “allelic variants,” andrefer to one of several alternate forms of a gene occupying a givenlocus on a chromosome of an organism. (Genes II, Lewin, B., ed., JohnWiley & Sons, New York (1985).) These allelic variants can vary ateither the polynucleotide and/or polypeptide level. Alternatively,non-naturally occurring variants may be produced by mutagenesistechniques or by direct synthesis.

[0282] Using known methods of protein engineering and recombinant DNAtechnology, variants may be generated to improve or alter thecharacteristics of the polypeptides of the present invention. Forinstance, one or more amino acids can be deleted from the N-terminus orC-terminus of the secreted protein without substantial loss ofbiological function. The authors of Ron et al., J. Biol. Chem. 268:2984-2988 (1993), reported variant KGF proteins having heparin bindingactivity even after deleting 3 8, or 27 amino-terminal amino acidresidues. Similarly, Interferon gamma exhibited up to ten times higheractivity after deleting 8-10 amino acid residues from the carboxyterminus of this protein. (Dobeli et al., J. Biotechnology 7:199-216(1988).)

[0283] Moreover, ample evidence demonstrates that variants often retaina biological activity similar to that of the naturally occurringprotein. For example, Gayle and coworkers (J. Biol. Chem 268:22105-22111(1993)) conducted extensive mutational analysis of human cytokine IL-1a.They used random mutagenesis to generate over 3,500 individual IL-1amutants that averaged 2.5 amino acid changes per variant over the entirelength of the molecule. Multiple mutations were examined at everypossible amino acid position. The investigators found that “[m]ost ofthe molecule could be altered with little effect on either [binding orbiological activity].” (See, Abstract.) In fact, only 23 unique aminoacid sequences, out of more than 3,500 nucleotide sequences examined,produced a protein that significantly differed in activity fromwild-type.

[0284] Furthermore, even if deleting one or more amino acids from theN-terminus or C-terminus of a polypeptide results in modification orloss of one or more biological functions, other biological activitiesmay still be retained. For example, the ability of a deletion variant toinduce and/or to bind antibodies which recognize the secreted form willlikely be retained when less than the majority of the residues of thesecreted form are removed from the N-terminus or C-terminus. Whether aparticular polypeptide lacking N- or C-terminal residues of a proteinretains such immunogenic activities can readily be determined by routinemethods described herein and otherwise known in the art.

[0285] Thus, the invention further includes polypeptide variants whichshow substantial biological activity. Such variants include deletions,insertions, inversions, repeats, and substitutions selected according togeneral rules known in the art so as have little effect on activity. Forexample, guidance concerning how to make phenotypically silent aminoacid substitutions is provided in Bowie, J. U. et al., Science247:1306-1310 (1990), wherein the authors indicate that there are twomain strategies for studying the tolerance of an amino acid sequence tochange.

[0286] The first strategy exploits the tolerance of amino acidsubstitutions by natural selection during the process of evolution. Bycomparing amino acid sequences in different species, conserved aminoacids can be identified. These conserved amino acids are likelyimportant for protein function. In contrast, the amino acid positionswhere substitutions have been tolerated by natural selection indicatesthat these positions are not critical for protein function. Thus,positions tolerating amino acid substitution could be modified whilestill maintaining biological activity of the protein.

[0287] The second strategy uses genetic engineering to introduce aminoacid changes at specific positions of a cloned gene to identify regionscritical for protein function. For example, site directed mutagenesis oralanine-scanning mutagenesis (introduction of single alanine mutationsat every residue in the molecule) can be used. (Cunningham and Wells,Science 244:1081-1085 (1989).) The resulting mutant molecules can thenbe tested for biological activity.

[0288] As the authors state, these two strategies have revealed thatproteins are surprisingly tolerant of amino acid substitutions. Theauthors further indicate which amino acid changes are likely to bepermissive at certain amino acid positions in the protein. For example,most buried (within the tertiary structure of the protein) amino acidresidues require nonpolar side chains, whereas few features of surfaceside chains are generally conserved. Moreover, tolerated conservativeamino acid substitutions involve replacement of the aliphatic orhydrophobic amino acids Ala, Val, Leu and Ile; replacement of thehydroxyl residues Ser and Thr; replacement of the acidic residues Aspand Glu; replacement of the amide residues Asn and Gln, replacement ofthe basic residues Lys, Arg, and His; replacement of the aromaticresidues Phe, Tyr, and Trp, and replacement of the small-sized aminoacids Ala, Ser, Thr, Met, and Gly.

[0289] Besides conservative amino acid substitution, variants of thepresent invention include (i) substitutions with one or more of thenon-conserved amino acid residues, where the substituted amino acidresidues may or may not be one encoded by the genetic code, or (ii)substitution with one or more of amino acid residues having asubstituent group, or (iii) fusion of the mature polypeptide withanother compound, such as a compound to increase the stability and/orsolubility of the polypeptide (for example, polyethylene glycol), or(iv) fusion of the polypeptide with additional amino acids, such as anIgG Fc fusion region peptide, or leader or secretory sequence, or asequence facilitating purification. Such variant polypeptides are deemedto be within the scope of those skilled in the art from the teachingsherein.

[0290] For example, polypeptide variants containing amino acidsubstitutions of charged amino acids with other charged or neutral aminoacids may produce proteins with improved characteristics, such as lessaggregation. Aggregation of pharmaceutical formulations both reducesactivity and increases clearance due to the aggregate's immunogenicactivity. (Pinckard et al., Clin. Exp. Immunol. 2:331-340 (1967);Robbins et al., Diabetes 36:838-845 (1987); Cleland et al., Crit. Rev.Therapeutic Drug Carrier Systems 10:307-377 (1993).)

[0291] Polynucleotide and Polypeptide Fragments

[0292] In the present invention, a “polynucleotide fragment” refers to ashort polynucleotide having a nucleic acid sequence contained in thedeposited clone or shown in SEQ ID NO:X. The short nucleotide fragmentsare preferably at least about 15 nt, and more preferably at least about20 nt, still more preferably at least about 30 nt, and even morepreferably, at least about 40 nt in length. A fragment “at least 20 ntin length,” for example, is intended to include 20 or more contiguousbases from the cDNA sequence contained in the deposited clone or thenucleotide sequence shown in SEQ ID NO:X. These nucleotide fragments areuseful as diagnostic probes and primers as discussed herein. Of course,larger fragments (e.g., 50, 150, 500, 600, 2000 nucleotides) arepreferred.

[0293] Moreover, representative examples of polynucleotide fragments ofthe invention, include, for example, fragments having a sequence fromabout nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250,251-300, 301-350, 351-400, 401-450, 451-500, 501-550, 551-600, 651-700,701-750, 751-800, 800-850, 851-900, 901-950, 951-1000, 1001-1050,1051-1100, 1101-1150, 1151-1200, 1201-1250, 1251-1300, 1301-1350,1351-1400, 1401-1450, 1451-1500, 1501-1550, 1551-1600, 1601-1650,1651-1700, 1701-1750, 1751-1800, 1801-1850, 1851-1900, 1901-1950,1951-2000, or 2001 to the end of SEQ ID NO:X or the cDNA contained inthe deposited clone. In this context “about” includes the particularlyrecited ranges, larger or smaller by several (5, 4, 3, 2, or 1)nucleotides, at either terminus or at both termini. Preferably, thesefragments encode a polypeptide which has biological activity. Morepreferably, these polynucleotides can be used as probes or primers asdiscussed herein.

[0294] In the present invention, a “polypeptide fragment” refers to ashort amino acid sequence contained in SEQ ID NO:Y or encoded by thecDNA contained in the deposited clone. Protein fragments may be“free-standing,” or comprised within a larger polypeptide of which thefragment forms a part or region, most preferably as a single continuousregion. Representative examples of polypeptide fragments of theinvention, include, for example, fragments from about amino acid number1-20, 21-40, 41-60, 61-80, 81-100, 102-120, 121-140, 141-160, or 161 tothe end of the coding region. Moreover, polypeptide fragments can beabout 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, or 150amino acids in length. In this context “about” includes the particularlyrecited ranges, larger or smaller by several (5, 4, 3, 2, or 1) aminoacids, at either extreme or at both extremes.

[0295] Preferred polypeptide fragments include the secreted protein aswell as the mature form. Further preferred polypeptide fragments includethe secreted protein or the mature form having a continuous series ofdeleted residues from the amino or the carboxy terminus, or both. Forexample, any number of amino acids, ranging from 1-60, can be deletedfrom the amino terminus of either the secreted polypeptide or the matureform. Similarly, any number of amino acids, ranging from 1-30, can bedeleted from the carboxy terminus of the secreted protein or matureform. Furthermore, any combination of the above amino and carboxyterminus deletions are preferred. Similarly, polynucleotide fragmentsencoding these polypeptide fragments are also preferred.

[0296] Also preferred are polypeptide and polynucleotide fragmentscharacterized by structural or functional domains, such as fragmentsthat comprise alpha-helix and alpha-helix forming regions, beta-sheetand beta-sheet-forming regions, turn and turn-forming regions, coil andcoil-forming regions, hydrophilic regions, hydrophobic regions, alphaamphipathic regions, beta amphipathic regions, flexible regions,surface-forming regions, substrate binding region, and high antigenicindex regions. Polypeptide fragments of SEQ ID NO:Y falling withinconserved domains are specifically contemplated by the presentinvention. Moreover, polynucleotide fragments encoding these domains arealso contemplated.

[0297] Other preferred fragments are biologically active fragments.Biologically active fragments are those exhibiting activity similar, butnot necessarily identical, to an activity of the polypeptide of thepresent invention. The biological activity of the fragments may includean improved desired activity, or a decreased undesirable activity.

[0298] Epitopes & Antibodies

[0299] In the present invention, “epitopes” refer to polypeptidefragments having antigenic or immunogenic activity in an animal,especially in a human. A preferred embodiment of the present inventionrelates to a polypeptide fragment comprising an epitope, as well as thepolynucleotide encoding this fragment. A region of a protein molecule towhich an antibody can bind is defined as an “antigenic epitope.” Incontrast, an “inmmunogenic epitope” is defined as a part of a proteinthat elicits an antibody response. (See, for instance, Geysen et al.,Proc. Natl. Acad. Sci. USA 8 1:3998-4002 (1983).)

[0300] Fragments which function as epitopes may be produced by anyconventional means. (See, e.g., Houghten, R. A., Proc. Natl. Acad. Sci.USA 82:5131-5135 (1985) further described in U.S. Pat. No. 4,631,211.)

[0301] In the present invention, antigenic epitopes preferably contain asequence of at least seven, more preferably at least nine, and mostpreferably between about 15 to about 30 amino acids. Antigenic epitopesare useful to raise antibodies, including monoclonal antibodies, thatspecifically bind the epitope. (See, for instance, Wilson et al., Cell37:767-778 (1984); Sutcliffe, J. G. et al., Science 219:660-666 (1983).)

[0302] Similarly, immunogenic epitopes can be used to induce antibodiesaccording to methods well known in the art. (See, for instance,Sutcliffe et al., supra; Wilson et al., supra; Chow, M. et al., Proc.Natl. Acad. Sci. USA 82:910-914; and Bittle, F. J. et al., J. Gen.Virol. 66:2347-2354 (1985).) A preferred immunogenic epitope includesthe secreted protein. The immunogenic epitopes may be presented togetherwith a carrier protein, such as an albumin, to an animal system (such asrabbit or mouse) or, if it is long enough (at least about 25 aminoacids), without a carrier. However, immunogenic epitopes comprising asfew as 8 to 10 amino acids have been shown to be sufficient to raiseantibodies capable of binding to, at the very least, linear epitopes ina denatured polypeptide (e.g., in Western blotting.)

[0303] As used herein, the term “antibody” (Ab) or “monoclonal antibody”(Mab) is meant to include intact molecules as well as antibody fragments(such as, for example, Fab and F(ab′)2 fragments) which are capable ofspecifically binding to protein. Fab and F(ab′)2 fragments lack the Fcfragment of intact antibody, clear more rapidly from the circulation,and may have less non-specific tissue binding than an intact antibody.(Wahl et al., J. Nucl. Med. 24:316-325 (1983).) Thus, these fragmentsare preferred, as well as the products of a FAB or other immunoglobulinexpression library. Moreover, antibodies of the present inventioninclude chimeric, single chain, and humanized antibodies.

[0304] Fusion Proteins

[0305] Any polypeptide of the present invention can be used to generatefusion proteins. For example, the polypeptide of the present invention,when fused to a second protein, can be used as an antigenic tag.Antibodies raised against the polypeptide of the present invention canbe used to indirectly detect the second protein by binding to thepolypeptide. Moreover, because secreted proteins target cellularlocations based on trafficking signals, the polypeptides of the presentinvention can be used as targeting molecules once fused to otherproteins.

[0306] Examples of domains that can be fused to polypeptides of thepresent invention include not only heterologous signal sequences, butalso other heterologous functional regions. The fusion does notnecessarily need to be direct, but may occur through linker sequences.

[0307] Moreover, fusion proteins may also be engineered to improvecharacteristics of the polypeptide of the present invention. Forinstance, a region of additional amino acids, particularly charged aminoacids, may be added to the N-terminus of the polypeptide to improvestability and persistence during purification from the host cell orsubsequent handling and storage. Also, peptide moieties may be added tothe polypeptide to facilitate purification. Such regions may be removedprior to final preparation of the polypeptide. The addition of peptidemoieties to facilitate handling of polypeptides are familiar and routinetechniques in the art.

[0308] Moreover, polypeptides of the present invention, includingfragments, and specifically epitopes, can be combined with parts of theconstant domain of immunoglobulins (IgG), resulting in chimericpolypeptides. These fusion proteins facilitate purification and show anincreased half-life in vivo. One reported example describes chimericproteins consisting of the first two domains of the humanCD4-polypeptide and various domains of the constant regions of the heavyor light chains of mammalian immunoglobulins. (EP A 394,827; Trauneckeret al., Nature 331:84-86 (1988).) Fusion proteins havingdisulfide-linked dimeric structures (due to the IgG) can also be moreefficient in binding and neutralizing other molecules, than themonomeric secreted protein or protein fragment alone. (Fountoulakis etal., J. Biochem. 270:3958-3964 (1995).)

[0309] Similarly, EP-A-O 464 533 (Canadian counterpart 2045869)discloses fusion proteins comprising various portions of constant regionof immunoglobulin molecules together with another human protein or partthereof. In many cases, the Fc part in a fusion protein is beneficial intherapy and diagnosis, and thus can result in, for example, improvedpharmacokinetic properties. (EP-A 0232 262.) Alternatively, deleting theFc part after the fusion protein has been expressed, detected, andpurified, would be desired. For example, the Fc portion may hindertherapy and diagnosis if the fusion protein is used as an antigen forimmunizations. In drug discovery, for example, human proteins, such ashIL-5, have been fused with Fc portions for the purpose ofhigh-throughput screening assays to identify antagonists of hIL-5. (See,D. Bennett et al., J. Molecular Recognition 8:52-58 (1995); K. Johansonet al., J. Biol. Chem. 270:9459-9471 (1995).)

[0310] Moreover, the polypeptides of the present invention can be fusedto marker sequences, such as a peptide which facilitates purification ofthe fused polypeptide. In preferred embodiments, the marker amino acidsequence is a hexa-histidine peptide, such as the tag provided in a pQEvector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311),among others, many of which are commercially available. As described inGentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), forinstance, hexa-histidine provides for convenient purification of thefusion protein. Another peptide tag useful for purification, the “HA”tag, corresponds to an epitope derived from the influenza hemagglutininprotein. (Wilson et al., Cell 37:767 (1984).)

[0311] Thus, any of these above fusions can be engineered using thepolynucleotides or the polypeptides of the present invention.

[0312] Vectors, Host Cells, and Protein Production

[0313] The present invention also relates to vectors containing thepolynucleotide of the present invention, host cells, and the productionof polypeptides by recombinant techniques. The vector may be, forexample, a phage, plasmid, viral, or retroviral vector. Retroviralvectors may be replication competent or replication defective. In thelatter case, viral propagation generally will occur only incomplementing host cells.

[0314] The polynucleotides may be joined to a vector containing aselectable marker for propagation in a host. Generally, a plasmid vectoris introduced in a precipitate, such as a calcium phosphate precipitate,or in a complex with a charged lipid. If the vector is a virus, it maybe packaged in vitro using an appropriate packaging cell line and thentransduced into host cells.

[0315] The polynucleotide insert should be operatively linked to anappropriate promoter, such as the phage lambda PL promoter, the E. colilac, trp, phoA and tac promoters, the SV40 early and late promoters andpromoters of retroviral LTRs, to name a few. Other suitable promoterswill be known to the skilled artisan. The expression constructs willfurther contain sites for transcription initiation, termination, and, inthe transcribed region, a ribosome binding site for translation. Thecoding portion of the transcripts expressed by the constructs willpreferably include a translation initiating codon at the beginning and atermination codon (UAA, UGA or UAG) appropriately positioned at the endof the polypeptide to be translated.

[0316] As indicated, the expression vectors will preferably include atleast one selectable marker. Such markers include dihydrofolatereductase, G418 or neomycin resistance for eukaryotic cell culture andtetracycline, kanamycin or ampicillin resistance genes for culturing inE. coli and other bacteria. Representative examples of appropriate hostsinclude, but are not limited to, bacterial cells, such as E. coli,Streptomyces and Salmonella typhimurium cells; fungal cells, such asyeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9cells; animal cells such as CHO, COS, 293, and Bowes melanoma cells; andplant cells. Appropriate culture mediums and conditions for theabove-described host cells are known in the art.

[0317] Among vectors preferred for use in bacteria include pQE70, pQE60and pQE-9, available from QIAGEN, Inc.; pBluescript vectors, Phagescriptvectors, pNH8A, pNH16a, pNH18A, pNH46A, available from StratageneCloning Systems, Inc.; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5available from Pharmacia Biotech, Inc. Among preferred eukaryoticvectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available fromStratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia.Other suitable vectors will be readily apparent to the skilled artisan.

[0318] Introduction of the construct into the host cell can be effectedby calcium phosphate transfection, DEAE-dextran mediated transfection,cationic lipid-mediated transfection, electroporation, transduction,infection, or other methods. Such methods are described in many standardlaboratory manuals, such as Davis et al., Basic Methods In MolecularBiology (1986). It is specifically contemplated that the polypeptides ofthe present invention may in fact be expressed by a host cell lacking arecombinant vector.

[0319] A polypeptide of this invention can be recovered and purifiedfrom recombinant cell cultures by well-known methods including ammoniumsulfate or ethanol precipitation, acid extraction, anion or cationexchange chromatography, phosphocellulose chromatography, hydrophobicinteraction chromatography, affinity chromatography, hydroxylapatitechromatography and lectin chromatography. Most preferably, highperformance liquid chromatography (“HPLC”) is employed for purification.

[0320] Polypeptides of the present invention, and preferably thesecreted form, can also be recovered from: products purified fromnatural sources, including bodily fluids, tissues and cells, whetherdirectly isolated or cultured; products of chemical syntheticprocedures; and products produced by recombinant techniques from aprokaryotic or eukaryotic host, including, for example, bacterial,yeast, higher plant, insect, and mammalian cells. Depending upon thehost employed in a recombinant production procedure, the polypeptides ofthe present invention may be glycosylated or may be non-glycosylated. Inaddition, polypeptides of the invention may also include an initialmodified methionine residue, in some cases as a result of host-mediatedprocesses. Thus, it is well known in the art that the N-terminalmethionine encoded by the translation initiation codon generally isremoved with high efficiency from any protein after translation in alleukaryotic cells. While the N-terminal methionine on most proteins alsois efficiently removed in most prokaryotes, for some proteins, thisprokaryotic removal process is inefficient, depending on the nature ofthe amino acid to which the N-terminal methionine is covalently linked.

[0321] Uses of the Polynucleotides

[0322] Each of the polynucleotides identified herein can be used innumerous ways as reagents. The following description should beconsidered exemplary and utilizes known techniques.

[0323] The polynucleotides of the present invention are useful forchromosome identification. There exists an ongoing need to identify newchromosome markers, since few chromosome marking reagents, based onactual sequence data (repeat polymorphisms), are presently available.Each polynucleotide of the present invention can be used as a chromosomemarker.

[0324] Briefly, sequences can be mapped to chromosomes by preparing PCRprimers (preferably 15-25 bp) from the sequences shown in SEQ ID NO:X.Primers can be selected using computer analysis so that primers do notspan more than one predicted exon in the genomic DNA. These primers arethen used for PCR screening of somatic cell hybrids containingindividual human chromosomes. Only those hybrids containing the humangene corresponding to the SEQ ID NO:X will yield an amplified fragment.

[0325] Similarly, somatic hybrids provide a rapid method of PCR mappingthe polynucleotides to particular chromosomes. Three or more clones canbe assigned per day using a single thermal cycler. Moreover,sublocalization of the polynucleotides can be achieved with panels ofspecific chromosome fragments. Other gene mapping strategies that can beused include in situ hybridization, prescreening with labeledflow-sorted chromosomes, and preselection by hybridization to constructchromosome specific-cDNA libraries.

[0326] Precise chromosomal location of the polynucleotides can also beachieved using fluorescence in situ hybridization (FISH) of a metaphasechromosomal spread. This technique uses polynucleotides as short as 500or 600 bases; however, polynucleotides 2,000-4,000 bp are preferred. Fora review of this technique, see Verma et al., “Human Chromosomes: aManual of Basic Techniques,” Pergamon Press, New York (1988).

[0327] For chromosome mapping, the polynucleotides can be usedindividually (to mark a single chromosome or a single site on thatchromosome) or in panels (for marking multiple sites and/or multiplechromosomes). Preferred polynucleotides correspond to the noncodingregions of the cDNAs because the coding sequences are more likelyconserved within gene families, thus increasing the chance of crosshybridization during chromosomal mapping.

[0328] Once a polynucleotide has been mapped to a precise chromosomallocation, the physical position of the polynucleotide can be used inlinkage analysis. Linkage analysis establishes coinheritance between achromosomal location and presentation of a particular disease. (Diseasemapping data are found, for example, in V. McKusick, MendelianInheritance in Man (available on line through Johns Hopkins UniversityWelch Medical Library) .) Assuming 1 megabase mapping resolution and onegene per 20 kb, a cDNA precisely localized to a chromosomal regionassociated with the disease could be one of 50-500 potential causativegenes.

[0329] Thus, once coinheritance is established, differences in thepolynucleotide and the corresponding gene between affected andunaffected individuals can be examined. First, visible structuralalterations in the chromosomes, such as deletions or translocations, areexamined in chromosome spreads or by PCR. If no structural alterationsexist, the presence of point mutations are ascertained. Mutationsobserved in some or all affected individuals, but not in normalindividuals, indicates that the mutation may cause the disease. However,complete sequencing of the polypeptide and the corresponding gene fromseveral normal individuals is required to distinguish the mutation froma polymorphism. If a new polymorphism is identified, this polymorphicpolypeptide can be used for further linkage analysis.

[0330] Furthermore, increased or decreased expression of the gene inaffected individuals as compared to unaffected individuals can beassessed using polynucleotides of the present invention. Any of thesealterations (altered expression, chromosomal rearrangement, or mutation)can be used as a diagnostic or prognostic marker.

[0331] In addition to the foregoing, a polynucleotide can be used tocontrol gene expression through triple helix formation or antisense DNAor RNA. Both methods rely on binding of the polynucleotide to DNA orRNA. For these techniques, preferred polynucleotides are usually 20 to40 bases in length and complementary to either the region of the geneinvolved in transcription (triple helix—see Lee et al., Nucl. Acids Res.6:3073 (1979); Cooney et al., Science 241:456 (1988); and Dervan et al.,Science 251:1360 (1991)) or to the mRNA itself (antisense—Okano, J.Neurochem. 56:560 (1991); Oligodeoxy-nucleotides as Antisense Inhibitorsof Gene Expression, CRC Press, Boca Raton, Fla. (1988).) Triple helixformation optimally results in a shut-off of RNA transcription from DNA,while antisense RNA hybridization blocks translation of an mRNA moleculeinto polypeptide. Both techniques are effective in model systems, andthe information disclosed herein can be used to design antisense ortriple helix polynucleotides in an effort to treat disease.

[0332] Polynucleotides of the present invention are also useful in genetherapy. One goal of gene therapy is to insert a normal gene into anorganism having a defective gene, in an effort to correct the geneticdefect. The polynucleotides disclosed in the present invention offer ameans of targeting such genetic defects in a highly accurate manner.Another goal is to insert a new gene that was not present in the hostgenome, thereby producing a new trait in the host cell.

[0333] The polynucleotides are also useful for identifying individualsfrom minute biological samples. The United States military, for example,is considering the use of restriction fragment length polymorphism(RFLP) for identification of its personnel. In this technique, anindividual's genomic DNA is digested with one or more restrictionenzymes, and probed on a Southern blot to yield unique bands foridentifying personnel. This method does not suffer from the currentlimitations of “Dog Tags” which can be lost, switched, or stolen, makingpositive identification difficult. The polynucleotides of the presentinvention can be used as additional DNA markers for RFLP.

[0334] The polynucleotides of the present invention can also be used asan alternative to RFLP, by determining the actual base-by-base DNAsequence of selected portions of an individual's genome. These sequencescan be used to prepare PCR primers for amplifying and isolating suchselected DNA, which can then be sequenced. Using this technique,individuals can be identified because each individual will have a uniqueset of DNA sequences. Once an unique ID database is established for anindividual, positive identification of that individual, living or dead,can be made from extremely small tissue samples.

[0335] Forensic biology also benefits from using DNA-basedidentification techniques as disclosed herein. DNA sequences taken fromvery small biological samples such as tissues, e.g., hair or skin, orbody fluids, e.g., blood, saliva, semen, etc., can be amplified usingPCR. In one prior art technique, gene sequences amplified frompolymorphic loci, such as DQa class II HLA gene, are used in forensicbiology to identify individuals. (Erlich, H., PCR Technology, Freemanand Co. (1992).) Once these specific polymorphic loci are amplified,they are digested with one or more restriction enzymes, yielding anidentifying set of bands on a Southern blot probed with DNAcorresponding to the DQa class II HLA gene. Similarly, polynucleotidesof the present invention can be used as polymorphic markers for forensicpurposes.

[0336] There is also a need for reagents capable of identifying thesource of a particular tissue. Such need arises, for example, inforensics when presented with tissue of unknown origin. Appropriatereagents can comprise, for example, DNA probes or primers specific toparticular tissue prepared from the sequences of the present invention.Panels of such reagents can identify tissue by species and/or by organtype. In a similar fashion, these reagents can be used to screen tissuecultures for contamination.

[0337] In the very least, the polynucleotides of the present inventioncan be used as molecular weight markers on Southern gels, as diagnosticprobes for the presence of a specific mRNA in a particular cell type, asa probe to “subtract-out” known sequences in the process of discoveringnovel polynucleotides, for selecting and making oligomers for attachmentto a “gene chip” or other support, to raise anti-DNA antibodies usingDNA immunization techniques, and as an antigen to elicit an immuneresponse.

[0338] Uses of the Polypeptides

[0339] Each of the polypeptides identified herein can be used innumerous ways. The following description should be considered exemplaryand utilizes known techniques.

[0340] A polypeptide of the present invention can be used to assayprotein levels in a biological sample using antibody-based techniques.For example, protein expression in tissues can be studied with classicalimmunohistological methods. (Jalkanen, M., et al., J. Cell. Biol.101:976-985 (1985); Jalkanen, M., et al., J. Cell . Biol. 105:3087-3096(1987).) Other antibody-based methods useful for detecting protein geneexpression include immunoassays, such as the enzyme linked immunosorbentassay (ELISA) and the radioimmunoassay (RIA). Suitable antibody assaylabels are known in the art and include enzyme labels, such as, glucoseoxidase, and radioisotopes, such as iodine (125I, 121I), carbon (14C),sulfur (35S), tritium (3H), indium (112In), and technetium (99mTc), andfluorescent labels, such as fluorescein and rhodamine, and biotin.

[0341] In addition to assaying secreted protein levels in a biologicalsample, proteins can also be detected in vivo by imaging. Antibodylabels or markers for in vivo imaging of protein include thosedetectable by X-radiography, NMR or ESR. For X-radiography, suitablelabels include radioisotopes such as barium or cesium, which emitdetectable radiation but are not overtly harmful to the subject.Suitable markers for NMR and ESR include those with a detectablecharacteristic spin, such as deuterium, which may be incorporated intothe antibody by labeling of nutrients for the relevant hybridoma.

[0342] A protein-specific antibody or antibody fragment which has beenlabeled with an appropriate detectable imaging moiety, such as aradioisotope (for example, 131I, 112In, 99mTc), a radio-opaquesubstance, or a material detectable by nuclear magnetic resonance, isintroduced (for example, parenterally, subcutaneously, orintraperitoneally) into the mammal. It will be understood in the artthat the size of the subject and the imaging system used will determinethe quantity of imaging moiety needed to produce diagnostic images. Inthe case of a radioisotope moiety, for a human subject, the quantity ofradioactivity injected will normally range from about 5 to 20millicuries of 99 mTc. The labeled antibody or antibody fragment willthen preferentially accumulate at the location of cells which containthe specific protein. In vivo tumor imaging is described in S. W.Burchiel et al., “Immunopharmacokinetics of Radiolabeled Antibodies andTheir Fragments.” (Chapter 13 in Tumor Imaging: The RadiochemicalDetection of Cancer, S. W. Burchiel and B. A. Rhodes, eds., MassonPublishing Inc. (1982).)

[0343] Thus, the invention provides a diagnostic method of a disorder,which involves (a) assaying the expression of a polypeptide of thepresent invention in cells or body fluid of an individual; (b) comparingthe level of gene expression with a standard gene expression level,whereby an increase or decrease in the assayed polypeptide geneexpression level compared to the standard expression level is indicativeof a disorder.

[0344] Moreover, polypeptides of the present invention can be used totreat disease. For example, patients can be administered a polypeptideof the present invention in an effort to replace absent or decreasedlevels of the polypeptide (e.g., insulin), to supplement absent ordecreased levels of a different polypeptide (e.g., hemoglobin S forhemoglobin B), to inhibit the activity of a polypeptide (e.g., anoncogene), to activate the activity of a polypeptide (e.g., by bindingto a receptor), to reduce the activity of a membrane bound receptor bycompeting with it for free ligand (e.g., soluble TNF receptors used inreducing inflammation), or to bring about a desired response (e.g.,blood vessel growth).

[0345] Similarly, antibodies directed to a polypeptide of the presentinvention can also be used to treat disease. For example, administrationof an antibody directed to a polypeptide of the present invention canbind and reduce overproduction of the polypeptide. Similarly,administration of an antibody can activate the polypeptide, such as bybinding to a polypeptide bound to a membrane (receptor).

[0346] At the very least, the polypeptides of the present invention canbe used as molecular weight markers on SDS-PAGE gels or on molecularsieve gel filtration columns using methods well known to those of skillin the art. Polypeptides can also be used to raise antibodies, which inturn are used to measure protein expression from a recombinant cell, asa way of assessing transformation of the host cell. Moreover, thepolypeptides of the present invention can be used to test the followingbiological activities.

[0347] Biological Activities

[0348] The polynucleotides and polypeptides of the present invention canbe used in assays to test for one or more biological activities. Ifthese polynucleotides and polypeptides do exhibit activity in aparticular assay, it is likely that these molecules may be involved inthe diseases associated with the biological activity. Thus, thepolynucleotides and polypeptides could be used to treat the associateddisease.

[0349] Immune Activity

[0350] A polypeptide or polynucleotide of the present invention may beuseful in treating deficiencies or disorders of the immune system, byactivating or inhibiting the proliferation, differentiation, ormobilization (chemotaxis) of immune cells. Immune cells develop througha process called hematopoiesis, producing myeloid (platelets, red bloodcells, neutrophils, and macrophages) and lymphoid (B and T lymphocytes)cells from pluripotent stem cells. The etiology of these immunedeficiencies or disorders may be genetic, somatic, such as cancer orsome autoimmune disorders, acquired (e.g., by chemotherapy or toxins),or infectious. Moreover, a polynucleotide or polypeptide of the presentinvention can be used as a marker or detector of a particular immunesystem disease or disorder.

[0351] A polynucleotide or polypeptide of the present invention may beuseful in treating or detecting deficiencies or disorders ofhematopoietic cells. A polypeptide or polynucleotide of the presentinvention could be used to increase differentiation and proliferation ofhematopoietic cells, including the pluripotent stem cells, in an effortto treat those disorders associated with a decrease in certain (or many)types hematopoietic cells. Examples of immunologic deficiency syndromesinclude, but are not limited to: blood protein disorders (e.g.agammaglobulinemia, dysgammaglobulinemia), ataxia telangiectasia, commonvariable immunodeficiency, Digeorge Syndrome, HIV infection, HTLV-BLVinfection, leukocyte adhesion deficiency syndrome, lymphopenia,phagocyte bactericidal dysfunction, severe combined immunodeficiency(SCIDs), Wiskott-Aldrich Disorder, anemia, thrombocytopenia, orhemoglobinuria.

[0352] Moreover, a polypeptide or polynucleotide of the presentinvention could also be used to modulate hemostatic (the stopping ofbleeding) or thrombolytic activity (clot formation). For example, byincreasing hemostatic or thrombolytic activity, a polynucleotide orpolypeptide of the present invention could be used to treat bloodcoagulation disorders (e.g., afibrinogenemia, factor deficiencies),blood platelet disorders (e.g. thrombocytopenia), or wounds resultingfrom trauma, surgery, or other causes. Alternatively, a polynucleotideor polypeptide of the present invention that can decrease hemostatic orthrombolytic activity could be used to inhibit or dissolve clotting.These molecules could be important in the treatment of heart attacks(infarction), strokes, or scarring.

[0353] A polynucleotide or polypeptide of the present invention may alsobe useful in treating or detecting autoimmune disorders. Many autoimmunedisorders result from inappropriate recognition of self as foreignmaterial by immune cells. This inappropriate recognition results in animmune response leading to the destruction of the host tissue.Therefore, the administration of a polypeptide or polynucleotide of thepresent invention that inhibits an immune response, particularly theproliferation, differentiation, or chemotaxis of T-cells, may be aneffective therapy in preventing autoimmune disorders.

[0354] Examples of autoimmune disorders that can be treated or detectedby the present invention include, but are not limited to: Addison'sDisease, hemolytic anemia, antiphospholipid syndrome, rheumatoidarthritis, dermatitis, allergic encephalomyelitis, glomerulonephritis,Goodpasture's Syndrome, Graves' Disease, Multiple Sclerosis, MyastheniaGravis, Neuritis, Ophthalmia, Bullous Pemphigoid, Pemphigus,Polyendocrinopathies, Purpura, Reiter's Disease, Stiff-Man Syndrome,Autoimmune Thyroiditis, Systemic Lupus Erythematosus, AutoimmunePulmonary Inflammation, Guillain-Barre Syndrome, insulin dependentdiabetes mellitis, and autoimmune inflammatory eye disease.

[0355] Similarly, allergic reactions and conditions, such as asthma(particularly allergic asthma) or other respiratory problems, may alsobe treated by a polypeptide or polynucleotide of the present invention.Moreover, these molecules can be used to treat anaphylaxis,hypersensitivity to an antigenic molecule, or blood groupincompatibility.

[0356] A polynucleotide or polypeptide of the present invention may alsobe used to treat and/or prevent organ rejection or graft-versus-hostdisease (GVHD). Organ rejection occurs by host immune cell destructionof the transplanted tissue through an immune response. Similarly, animmune response is also involved in GVHD, but, in this case, the foreigntransplanted immune cells destroy the host tissues. The administrationof a polypeptide or polynucleotide of the present invention thatinhibits an immune response, particularly the proliferation,differentiation, or chemotaxis of T-cells, may be an effective therapyin preventing organ rejection or GVHD.

[0357] Similarly, a polypeptide or polynucleotide of the presentinvention may also be used to modulate inflammation. For example, thepolypeptide or polynucleotide may inhibit the proliferation anddifferentiation of cells involved in an inflammatory response. Thesemolecules can be used to treat inflammatory conditions, both chronic andacute conditions, including inflammation associated with infection(e.g., septic shock, sepsis, or systemic inflammatory response syndrome(SIRS)), ischemnia-reperfusion injury, endotoxin lethality, arthritis,complement-mediated hyperacute rejection, nephritis, cytokine orchemokine induced lung injury, inflammatory bowel disease, Crohn'sdisease, or resulting from over production of cytokines (e.g., TNF orIL- 1.)

[0358] Hyperproliferative Disorders

[0359] A polypeptide or polynucleotide can be used to treat or detecthyperproliferative disorders, including neoplasms. A polypeptide orpolynucleotide of the present invention may inhibit the proliferation ofthe disorder through direct or indirect interactions. Alternatively, apolypeptide or polynucleotide of the present invention may proliferateother cells which can inhibit the hyperproliferative disorder.

[0360] For example, by increasing an immune response, particularlyincreasing antigenic qualities of the hyperproliferative disorder or byproliferating, differentiating, or mobilizing T-cells,hyperproliferative disorders can be treated. This immune response may beincreased by either enhancing an existing immune response, or byinitiating a new immune response. Alternatively, decreasing an immuneresponse may also be a method of treating hyperproliferative disorders,such as a chemotherapeutic agent.

[0361] Examples of hyperproliferative disorders that can be treated ordetected by a polynucleotide or polypeptide of the present inventioninclude, but are not limited to neoplasms located in the: abdomen, bone,breast, digestive system, liver, pancreas, peritoneum, endocrine glands(adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid),eye, head and neck, nervous (central and peripheral), lymphatic system,pelvic, skin, soft tissue, spleen, thoracic, and urogenital.

[0362] Similarly, other hyperproliferative disorders can also be treatedor detected by a polynucleotide or polypeptide of the present invention.Examples of such hyperproliferative disorders include, but are notlimited to: hypergammaglobulinemia, lymphoproliferative disorders,paraproteinemias, purpura, sarcoidosis, Sezary Syndrome, Waldenstron'sMacroglobulinemia, Gaucher's Disease, histiocytosis, and any otherhyperproliferative disease, besides neoplasia, located in an organsystem listed above.

[0363] Infectious Disease

[0364] A polypeptide or polynucleotide of the present invention can beused to treat or detect infectious agents. For example, by increasingthe immune response, particularly increasing the proliferation anddifferentiation of B and/or T cells, infectious diseases may be treated.The immune response may be increased by either enhancing an existingimmune response, or by initiating a new immune response. Alternatively,the polypeptide or polynucleotide of the present invention may alsodirectly inhibit the infectious agent, without necessarily eliciting animmune response.

[0365] Viruses are one example of an infectious agent that can causedisease or symptoms that can be treated or detected by a polynucleotideor polypeptide of the present invention. Examples of viruses, include,but are not limited to the following DNA and RNA viral families:Arbovirus, Adenoviridae, Arenaviridae, Arterivirus, Birnaviridae,Bunyaviridae, Caliciviridae, Circoviridae, Coronaviridae, Flaviviridae,Hepadnaviridae (Hepatitis), Herpesviridae (such as, Cytomegalovirus,Herpes Simplex, Herpes Zoster), Mononegavirus (e.g., Paramyxoviridae,Morbillivirus, Rhabdoviridae), Orthomyxoviridae (e.g., Influenza),Papovaviridae, Parvoviridae, Picomaviridae, Poxviridae (such as Smallpoxor Vaccinia), Reoviridae (e.g., Rotavirus), Retroviridae (HTLV-I,HTLV-II, Lentivirus), and Togaviridae (e.g., Rubivirus). Viruses fallingwithin these families can cause a variety of diseases or symptoms,including, but not limited to: arthritis, bronchiollitis, encephalitis,eye infections (e.g., conjunctivitis. keratitis), chronic fatiguesyndrome, hepatitis (A, B, C, E, Chronic Active, Delta), meningitis,opportunistic infections (e.g., AIDS), pneumonia, Burkitt's Lymphoma,chickenpox, hemorrhagic fever, Measles, Mumps, Parainfluenza, Rabies,the common cold, Polio, leukemia, Rubella, sexually transmitteddiseases, skin diseases (e.g., Kaposi's, warts), and viremia. Apolypeptide or polynucleotide of the present invention can be used totreat or detect any of these symptoms or diseases.

[0366] Similarly, bacterial or fungal agents that can cause disease orsymptoms and that can be treated or detected by a polynucleotide orpolypeptide of the present invention include, but not limited to, thefollowing Gram-Negative and Gram-positive bacterial families and fungi:Actinomycetales (e.g., Corynebacterium, Mycobacterium, Norcardia),Aspergillosis, Bacillaceae (e.g., Anthrax, Clostridium), Bacteroidaceae,Blastomycosis, Bordetella, Borrelia, Brucellosis, Candidiasis,Campylobacter, Coccidioidomycosis, Cryptococcosis, Derrnatocycoses,Enterobacteriaceae (Klebsiella, Salmonella, Serratia, Yersinia),Erysipelothrix, Helicobacter, Legionellosis, Leptospirosis, Listeria,Mycoplasmatales, Neisseriaceae (e.g., Acinetobacter, Gonorrhea,Menigococcal), Pasteurellacea Infections (e.g., Actinobacillus,Heamophilus, Pasteurella), Pseudomonas, Rickettsiaceae, Chlamydiaceae,Syphilis, and Staphylococcal. These bacterial or fungal families cancause the following diseases or symptoms, including, but not limited to:bacteremia, endocarditis, eye infections (conjunctivitis, tuberculosis,uveitis), gingivitis, opportunistic infections (e.g., AIDS relatedinfections), paronychia, prosthesis-related infections, Reiter'sDisease, respiratory tract infections, such as Whooping Cough orEmpyema, sepsis, Lyme Disease, Cat-Scratch Disease, Dysentery,Paratyphoid Fever, food poisoning, Typhoid, pneumonia, Gonorrhea,meningitis, Chlamydia, Syphilis, Diphtheria, Leprosy, Paratuberculosis,Tuberculosis, Lupus, Botulism, gangrene, tetanus, impetigo, RheumaticFever, Scarlet Fever, sexually transmitted diseases, skin diseases(e.g., cellulitis, dermatocycoses), toxemia, urinary tract infections,wound infections. A polypeptide or polynucleotide of the presentinvention can be used to treat or detect any of these symptoms ordiseases.

[0367] Moreover, parasitic agents causing disease or symptoms that canbe treated or detected by a polynucleotide or polypeptide of the presentinvention include, but not liuited to, the following families:Arnebiasis, Babesiosis, Coccidiosis, Cryptosporidiosis, Dientamoebiasis,Dourine, Ectoparasitic, Giardiasis, Helminthiasis, Leishmaniasis,Theileriasis, Toxoplasmosis, Trypanosormiasis, and Trichomonas. Theseparasites can cause a variety of diseases or symptoms, including, butnot limited to: Scabies, Trombiculiasis, eye infections, intestinaldisease (e.g., dysentery, giardiasis), liver disease, lung disease,opportunistic infections (e.g., AIDS related), Malaria, pregnancycomplications, and toxoplasmosis. A polypeptide or polynucleotide of thepresent invention can be used to treat or detect any of these symptomsor diseases.

[0368] Preferably, treatment using a polypeptide or polynucleotide ofthe present invention could either be by administering an effectiveamount of a polypeptide to the patient, or by removing cells from thepatient, supplying the cells with a polynucleotide of the presentinvention, and returning the engineered cells to the patient (ex vivotherapy). Moreover, the polypeptide or polynucleotide of the presentinvention can be used as an antigen in a vaccine to raise an immuneresponse against infectious disease.

[0369] Regeneration

[0370] A polynucleotide or polypeptide of the present invention can beused to differentiate, proliferate, and attract cells, leading to theregeneration of tissues. (See, Science 276:59-87 (1997).) Theregeneration of tissues could be used to repair, replace, or protecttissue damaged by congenital defects, trauma (wounds, burns, incisions,or ulcers), age, disease (e.g. osteoporosis, osteocarthritis,periodontal disease, liver failure), surgery, including cosmetic plasticsurgery, fibrosis, reperfusion injury, or systemic cytokine damage.

[0371] Tissues that could be regenerated using the present inventioninclude organs (e.g., pancreas, liver, intestine, kidney, skin,endothelium), muscle (smooth, skeletal or cardiac), vascular (includingvascular endothelium), nervous, hematopoietic, and skeletal (bone,cartilage, tendon, and ligament) tissue. Preferably, regeneration occurswithout or decreased scarring. Regeneration also may includeangiogenesis.

[0372] Moreover, a polynucleotide or polypeptide of the presentinvention may increase regeneration of tissues difficult to heal. Forexample, increased tendon/ligament regeneration would quicken recoverytime after damage. A polynucleotide or polypeptide of the presentinvention could also be used prophylactically in an effort to avoiddamage. Specific diseases that could be treated include of tendinitis,carpal tunnel syndrome, and other tendon or ligament defects. A furtherexample of tissue regeneration of non-healing wounds includes pressureulcers, ulcers associated with vascular insufficiency, surgical, andtraumatic wounds.

[0373] Similarly, nerve and brain tissue could also be regenerated byusing a polynucleotide or polypeptide of the present invention toproliferate and differentiate nerve cells. Diseases that could betreated using this method include central and peripheral nervous systemdiseases, neuropathies, or mechanical and traumatic disorders (e.g.,spinal cord disorders, head trauma, cerebrovascular disease, and stoke).Specifically, diseases associated with peripheral nerve injuries,peripheral neuropathy (e.g., resulting from chemotherapy or othermedical therapies), localized neuropathies, and central nervous systemdiseases (e.g., Alzheimer's disease, Parkinson's disease, Huntington'sdisease, amyotrophic lateral sclerosis, and Shy-Drager syndrome), couldall be treated using the polynucleotide or polypeptide of the presentinvention.

[0374] Chemotaxis

[0375] A polynucleotide or polypeptide of the present invention may havechemotaxis activity. A chemotaxic molecule attracts or mobilizes cells(e.g., monocytes, fibroblasts, neutrophils, T-cells, mast cells,eosinophils, epithelial and/or endothelial cells) to a particular sitein the body, such as inflammation, infection, or site ofhyperproliferation. The mobilized cells can then fight off and/or healthe particular trauma or abnormality.

[0376] A polynucleotide or polypeptide of the present invention mayincrease chemotaxic activity of particular cells. These chemotacticmolecules can then be used to treat inflammation, infection,hyperproliferative disorders, or any immune system disorder byincreasing the number of cells targeted to a particular location in thebody. For example, chemotaxic molecules can be used to treat wounds andother trauma to tissues by attracting immune cells to the injuredlocation. Chemotactic molecules of the present invention can alsoattract fibroblasts, which can be used to treat wounds.

[0377] It is also contemplated that a polynucleotide or polypeptide ofthe present invention may inhibit chemotactic activity. These moleculescould also be used to treat disorders. Thus, a polynucleotide orpolypeptide of the present invention could be used as an inhibitor ofchemotaxis.

[0378] Binding Activity

[0379] A polypeptide of the present invention may be used to screen formolecules that bind to the polypeptide or for molecules to which thepolypeptide binds. The binding of the polypeptide and the molecule mayactivate (agonist), increase, inhibit (antagonist), or decrease activityof the polypeptide or the molecule bound. Examples of such moleculesinclude antibodies, oligonucleotides, proteins (e.g., receptors),orsmall molecules.

[0380] Preferably, the molecule is closely related to the natural ligandof the polypeptide, e.g., a fragment of the ligand, or a naturalsubstrate, a ligand, a structural or functional mimetic. (See, Coliganet al., Current Protocols in Immunology 1(2):Chapter 5 (1991).)Similarly, the molecule can be closely related to the natural receptorto which the polypeptide binds, or at least, a fragment of the receptorcapable of being bound by the polypeptide (e.g., active site). In eithercase, the molecule can be rationally designed using known techniques.

[0381] Preferably, the screening for these molecules involves producingappropriate cells which express the polypeptide, either as a secretedprotein or on the cell membrane. Preferred cells include cells frommammals, yeast, Drosophila, or E. coli. Cells expressing the polypeptide(or cell membrane containing the expressed polypeptide) are thenpreferably contacted with a test compound potentially containing themolecule to observe binding, stimulation, or inhibition of activity ofeither the polypeptide or the molecule.

[0382] The assay may simply test binding of a candidate compound to thepolypeptide, wherein binding is detected by a label, or in an assayinvolving competition with a labeled competitor. Further, the assay maytest whether the candidate compound results in a signal generated bybinding to the polypeptide.

[0383] Alternatively, the assay can be carried out using cell-freepreparations, polypeptide/molecule affixed to a solid support, chemicallibraries, or natural product mixtures. The assay may also simplycomprise the steps of mixing a candidate compound with a solutioncontaining a polypeptide, measuring polypeptidelmolecule activity orbinding, and comparing the polypeptide/molecule activity or binding to astandard.

[0384] Preferably, an ELISA assay can measure polypeptide level oractivity in a sample (e.g., biological sample) using a monoclonal orpolyclonal antibody. The antibody can measure polypeptide level oractivity by either binding, directly or indirectly, to the polypeptideor by competing with the polypeptide for a substrate.

[0385] All of these above assays can be used as diagnostic or prognosticmarkers. The molecules discovered using these assays can be used totreat disease or to bring about a particular result in a patient (e.g.,blood vessel growth) by activating or inhibiting thepolypeptide/molecule. Moreover, the assays can discover agents which mayinhibit or enhance the production of the polypeptide from suitablymanipulated cells or tissues.

[0386] Therefore, the invention includes a method of identifyingcompounds which bind to a polypeptide of the invention comprising thesteps of: (a) incubating a candidate binding compound with a polypeptideof the invention; and (b) determining if binding has occurred. Moreover,the invention includes a method of identifying agonists/antagonistscomprising the steps of: (a) incubating a candidate compound with apolypeptide of the invention, (b) assaying a biological activity , and(b) determining if a biological activity of the polypeptide has beenaltered.

[0387] Other Activities

[0388] A polypeptide or polynucleotide of the present invention may alsoincrease or decrease the differentiation or proliferation of embryonicstem cells, besides, as discussed above, hematopoietic lineage.

[0389] A polypeptide or polynucleotide of the present invention may alsobe used to modulate mammalian characteristics, such as body height,weight, hair color, eye color, skin, percentage of adipose tissue,pigmentation, size, and shape (e.g., cosmetic surgery). Similarly, apolypeptide or polynucleotide of the present invention may be used tomodulate mammalian metabolism affecting catabolism, anabolism,processing, utilization, and storage of energy.

[0390] A polypeptide or polynucleotide of the present invention may beused to change a mammal's mental state or physical state by influencingbiorhythms, caricadic rhythms, depression (including depressivedisorders), tendency for violence, tolerance for pain, reproductivecapabilities (preferably by Activin or Inhibin-like activity), hormonalor endocrine levels, appetite, libido, memory, stress, or othercognitive qualities.

[0391] A polypeptide or polynucleoeide of the present invention may alsobe used as a food additive or preservative, such as to increase ordecrease storage capabilities, fat content, lipid, protein,carbohydrate, vitamins, minerals, cofactors or other nutritionalcomponents.

[0392] Other Preferred Embodiments

[0393] Other preferred embodiments of the claimed invention include anisolated nucleic acid molecule comprising a nucleotide sequence which isat least 95% identical to a sequence of at least about 50 contiguousnucleotides in the nucleotide sequence of SEQ ID NO:X wherein X is anyinteger as defined in Table 1.

[0394] Also preferred is a nucleic acid molecule wherein said sequenceof contiguous nucleotides is included in the nucleotide sequence of SEQID NO:X in the range of positions beginning with the nucleotide at aboutthe position of the 5′ Nucleotide of the Clone Sequence and ending withthe nucleotide at about the position of the 3′ Nucleotide of the CloneSequence as defined for SEQ ID NO-X in Table 1.

[0395] Also preferred is a nucleic acid molecule wherein said sequenceof contiguous nucleotides is included in the nucleotide sequence of SEQID NO:X in the range of positions beginning with the nucleotide at aboutthe position of the 5′ Nucleotide of the Start Codon and ending with thenucleotide at about the position of the 3′ Nucleotide of the CloneSequence as defined for SEQ ID NO:X in Table 1.

[0396] Similarly preferred is a nucleic acid molecule wherein saidsequence of contiguous nucleotides is included in the nucleotidesequence of SEQ ID NO:X in the range of positions beginning with thenucleotide at about the position of the 5′ Nucleotide of the First AminoAcid of the Signal Peptide and ending with the nucleotide at about theposition of the 3′ Nucleotide of the Clone Sequence as defined for SEQID NO:X in Table 1.

[0397] Also preferred is an isolated nucleic acid molecule comprising anucleotide sequence which is at least 95% identical to a sequence of atleast about 150 contiguous nucleotides in the nucleotide sequence of SEQID NO:X.

[0398] Further preferred is an isolated nucleic acid molecule comprisinga nucleotide sequence which is at least 95% identical to a sequence ofat least about 500 contiguous nucleotides in the nucleotide sequence ofSEQ ID NO:X.

[0399] A further preferred embodiment is a nucleic acid moleculecomprising a nucleotide sequence which is at least 95% identical to thenucleotide sequence of SEQ ID NO:X beginning with the nucleotide atabout the position of the 5′ Nucleotide of the First Amino Acid of theSignal Peptide and ending with the nucleotide at about the position ofthe 3′ Nucleotide of the Clone Sequence as defined for SEQ ID NO:X inTable 1.

[0400] A further preferred embodiment is an isolated nucleic acidmolecule comprising a nucleotide sequence which is at least 95%identical to the complete nucleotide sequence of SEQ ID NO:X.

[0401] Also preferred is an isolated nucleic acid molecule whichhybridizes under stringent hybridization conditions to a nucleic acidmolecule, wherein said nucleic acid molecule which hybridizes does nothybridize under stringent hybridization conditions to a nucleic acidmolecule having a nucleotide sequence consisting of only A residues orof only T residues.

[0402] Also preferred is a composition of matter comprising a DNAmolecule which comprises a human cDNA clone identified by a cDNA CloneIdentifier in Table 1, which DNA molecule is contained in the materialdeposited with the American Type Culture Collection and given the ATCCDeposit Number shown in Table 1 for said cDNA Clone Identifier.

[0403] Also preferred is an isolated nucleic acid molecule comprising anucleotide sequence which is at least 95% identical to a sequence of atleast 50 contiguous nucleotides in the nucleotide sequence of a humancDNA clone identified by a cDNA Clone Identifier in Table 1, which DNAmolecule is contained in the deposit given the ATCC Deposit Number shownin Table 1.

[0404] Also preferred is an isolated nucleic acid molecule, wherein saidsequence of at least 50 contiguous nucleotides is included in thenucleotide sequence of the complete open reading frame sequence encodedby said human cDNA clone.

[0405] Also preferred is an isolated nucleic acid molecule comprising anucleotide sequence which is at least 95% identical to sequence of atleast 150 contiguous nucleotides in the nucleotide sequence encoded bysaid human cDNA clone.

[0406] A further preferred embodiment is an isolated nucleic acidmolecule comprising a nucleotide sequence which is at least 95%identical to sequence of at least 500 contiguous nucleotides in thenucleotide sequence encoded by said human cDNA clone.

[0407] A further preferred embodiment is an isolated nucleic acidmolecule comprising a nucleotide sequence which is at least 95%identical to the complete nucleotide sequence encoded by said human cDNAclone.

[0408] A further preferred embodiment is a method for detecting in abiological sample a nucleic acid molecule comprising a nucleotidesequence which is at least 95% identical to a sequence of at least 50contiguous nucleotides in a sequence selected from the group consistingof: a nucleotide sequence of SEQ ID NO:X wherein X is any integer asdefined in Table 1; and a nucleotide sequence encoded by a human cDNAclone identified by a cDNA Clone Identifier in Table 1 and contained inthe deposit with the ATCC Deposit Number shown for said cDNA clone inTable 1; which method comprises a step of comparing a nucleotidesequence of at least one nucleic acid molecule in said sample with asequence selected from said group and determining whether the sequenceof said nucleic acid molecule in said sample is at least 95% identicalto said selected sequence.

[0409] Also preferred is the above method wherein said step of comparingsequences comprises determining the extent of nucleic acid hybridizationbetween nucleic acid molecules in said sample and a nucleic acidmolecule comprising said sequence selected from said group. Similarly,also preferred is the above method wherein said step of comparingsequences is performed by comparing the nucleotide sequence determinedfrom a nucleic acid molecule in said sample with said sequence selectedfrom said group. The nucleic acid molecules can comprise DNA moleculesor RNA molecules.

[0410] A further preferred embodiment is a method for identifying thespecies, tissue or cell type of a biological sample which methodcomprises a step of detecting nucleic acid molecules in said sample, ifany, comprising a nucleotide sequence that is at least 95% identical toa sequence of at least 50 contiguous nucleotides in a sequence selectedfrom the group consisting of: a nucleotide sequence of SEQ ID NO:Xwherein X is any integer as defined in Table 1; and a nucleotidesequence encoded by a human cDNA clone identified by a cDNA CloneIdentifier in Table 1 and contained in the deposit with the ATCC DepositNumber shown for said cDNA clone in Table 1.

[0411] The method for identifying the species, tissue or cell type of abiological sample can comprise a step of detecting nucleic acidmolecules comprising a nucleotide sequence in a panel of at least twonucleotide sequences, wherein at least one sequence in said panel is atleast 95% identical to a sequence of at least 50 contiguous nucleotidesin a sequence selected from said group.

[0412] Also preferred is a method for diagnosing in a subject apathological condition associated with abnormal structure or expressionof a gene encoding a secreted protein identified in Table 1, whichmethod comprises a step of detecting in a biological sample obtainedfrom said subject nucleic acid molecules, if any, comprising anucleotide sequence that is at least 95% identical to a sequence of atleast 50 contiguous nucleotides in a sequence selected from the groupconsisting of: a nucleotide sequence of SEQ ID NO:X wherein X is anyinteger as defined in Table 1; and a nucleotide sequence encoded by ahuman cDNA clone identified by a cDNA Clone Identifier in Table 1 andcontained in the deposit with the ATCC Deposit Number shown for saidcDNA clone-in Table 1.

[0413] The method for diagnosing a pathological condition can comprise astep of detecting nucleic acid molecules comprising a nucleotidesequence in a panel of at least two nucleotide sequences, wherein atleast one sequence in said panel is at least 95% identical to a sequenceof at least 50 contiguous nucleotides in a sequence selected from saidgroup.

[0414] Also preferred is a composition of matter comprising isolatednucleic acid molecules wherein the nucleotide sequences of said nucleicacid molecules comprise a panel of at least two nucleotide sequences,wherein at least one sequence in said panel is at least 95% identical toa sequence of at-least 50 contiguous nucleotides in a sequence selectedfrom the group consisting of: a nucleotide sequence of SEQ ID NO:Xwherein X is any integer as defined in Table 1; and a nucleotidesequence encoded by a human cDNA clone identified by a cDNA CloneIdentifier in Table 1 and contained in the deposit with the ATCC DepositNumber shown for said cDNA clone in Table 1. The nucleic acid moleculescan comprise DNA molecules or RNA molecules.

[0415] Also preferred is an isolated polypeptide comprising an aminoacid sequence at least 90% identical to a sequence of at least about 10contiguous amino acids in the amino acid sequence of SEQ ID NO:Y whereinY is any integer as defined in Table 1.

[0416] Also preferred is a polypeptide, wherein said sequence ofcontiguous amino acids is included in the amino acid sequence of SEQ IDNO:Y in the range of positions beginning with the residue at about theposition of the First Amino Acid of the Secreted Portion and ending withthe residue at about the Last Amino Acid of the Open Reading Frame asset forth for SEQ ID NO:Y in Table 1.

[0417] Also preferred is an isolated polypeptide comprising an aminoacid sequence at least 95% identical to a sequence of at least about 30contiguous amino acids in the amino acid sequence of SEQ ID NO:Y.

[0418] Further preferred is an isolated polypeptide comprising an aminoacid sequence at least 95% identical to a sequence of at least about 100contiguous amino acids in the amino acid sequence of SEQ ID NO:Y.

[0419] Further preferred is an isolated polypeptide comprising an aminoacid sequence at least 95% identical to the complete amino acid sequenceof SEQ ID NO:Y.

[0420] Further preferred is an isolated polypeptide comprising an aminoacid sequence at least 90% identical to a sequence of at least about 10contiguous amino acids in the complete amino acid sequence of a secretedprotein encoded by a human cDNA clone identified by a cDNA CloneIdentifier in Table 1 and contained in the deposit with the ATCC DepositNumber shown for said cDNA clone in Table 1.

[0421] Also preferred is a polypeptide wherein said sequence ofcontiguous amino acids is included in the amino acid sequence of asecreted portion of the secreted protein encoded by a human cDNA cloneidentified by a cDNA Clone Identifier in Table 1 and contained in thedeposit with the ATCC Deposit Number shown for said cDNA clone in Table1.

[0422] Also preferred is an isolated polypeptide comprising an aminoacid sequence at least 95% identical to a sequence of at least about 30contiguous amino acids in the amino acid sequence of the secretedportion of the protein encoded by a human cDNA clone identified by acDNA Clone Identifier in Table 1 and contained in the deposit with theATCC Deposit Number shown for said cDNA clone in Table 1.

[0423] Also preferred is an isolated polypeptide comprising an aminoacid sequence at least 95% identical to a sequence of at least about 100contiguous amino acids in the amino acid sequence of the secretedportion of the protein encoded by a human cDNA clone identified by acDNA Clone Identifier in Table 1 and contained in the deposit with theATCC Deposit Number shown for said cDNA clone in Table 1.

[0424] Also preferred is an isolated polypeptide comprising an aminoacid sequence at least 95% identical to the amino acid sequence of thesecreted portion of the protein encoded by a human cDNA clone identifiedby a cDNA Clone Identifier in Table 1 and contained in the deposit withthe ATCC Deposit Number shown for said cDNA clone in Table 1.

[0425] Further preferred is an isolated antibody which bindsspecifically to a polypeptide comprising an amino acid sequence that isat least 90% identical to a sequence of at least 10 contiguous aminoacids in a sequence selected from the group consisting of: an amino acidsequence of SEQ ID NO:Y wherein Y is any integer as defined in Table 1;and a complete amino acid sequence of a protein encoded by a human cDNAclone identified by a cDNA Clone Identifier in Table 1 and contained inthe deposit with the ATCC Deposit Number shown for said cDNA clone inTable 1.

[0426] Further preferred is a method for detecting in a biologicalsample a polypeptide comprising an amino acid sequence which is at least90% identical to a sequence of at least 10 contiguous amino acids in asequence selected from the group consisting of: an amino acid sequenceof SEQ ID NO:Y wherein Y is any integer as defined in Table 1;

[0427] and a complete amino acid sequence of a protein encoded by ahuman cDNA clone identified by a cDNA Clone Identifier in Table 1 andcontained in the deposit with the ATCC Deposit Number shown for saidcDNA clone in Table 1; which method comprises a step of comparing anamino acid sequence of at least one polypeptide molecule in said samplewith a sequence selected from said group and determining whether thesequence of said polypeptide molecule in said sample is at least 90%identical to said sequence of at least 10 contiguous amino acids.

[0428] Also preferred is the above method wherein said step of comparingan amino acid sequence of at least one polypeptide molecule in saidsample with a sequence selected from said group comprises determiningthe extent of specific binding of polypeptides in said sample to anantibody which binds specifically to a polypeptide comprising an aminoacid sequence that is at least 90% identical to a sequence of at least10 contiguous amino acids in a sequence selected from the groupconsisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is anyinteger as defined in Table 1;

[0429] and a complete amino acid sequence of a protein encoded by ahuman cDNA clone identified by a cDNA Clone Identifier in Table 1 andcontained in the deposit with the ATCC Deposit Number shown for saidcDNA clone in Table 1.

[0430] Also preferred is the above method wherein said step of comparingsequences is performed by comparing the amino acid sequence determinedfrom a polypeptide molecule in said sample with said sequence selectedfrom said group.

[0431] Also preferred is a method for identifying the species, tissue orcell type of a biological sample which method comprises a step ofdetecting polypeptide molecules in said sample, if any, comprising anamino acid sequence that is at least 90% identical to a sequence of atleast 10 contiguous amino acids in a sequence selected from the groupconsisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is anyinteger as defined in Table 1; and a complete amino acid sequence of asecreted protein encoded by a human cDNA clone identified by a cDNAClone Identifier in Table 1 and contained in the deposit with the ATCCDeposit Number shown for said cDNA clone in Table 1.

[0432] Also preferred is the above method for identifying the species,tissue or cell type of a biological sample, which method comprises astep of detecting polypeptide molecules comprising an amino acidsequence in a panel of at least two amino acid sequences, wherein atleast one sequence in said panel is at least 90% identical to a sequenceof at least 10 contiguous amino acids in a sequence selected from theabove group.

[0433] Also preferred is a method for diagnosing in a subject apathological condition associated with abnormal stricture or expressionof a gene encoding a secreted protein identified in Table 1, whichmethod comprises a step of detecting in a biological sample obtainedfrom said subject polypeptide molecules comprising an amino acidsequence in a panel of at least two amino acid sequences, wherein atleast one sequence in said panel is at least 90% identical to a sequenceof at least 10 contiguous amino acids in a sequence selected from thegroup consisting of: an amino acid sequence of SEQ ID NO:Y wherein Y isany integer as defined in Table 1; and a complete amino acid sequence ofa secreted protein encoded by a human cDNA clone identified by a cDNAClone Identifier in Table 1 and contained in the deposit with the ATCCDeposit Number shown for said cDNA clone in Table 1.

[0434] In any of these methods, the step of detecting said polypeptidemolecules includes using an antibody.

[0435] Also preferred is an isolated nucleic acid molecule comprising anucleotide sequence which is at least 95% identical to a nucleotidesequence encoding a polypeptide wherein said polypeptide comprises anamino acid sequence that is at least 90% identical to a sequence of atleast 10 contiguous amino acids in a sequence selected from the groupconsisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is anyinteger as defined in Table 1; and a complete amino acid sequence of asecreted protein encoded by a human cDNA clone identified by a cDNAClone Identifier in Table 1 and contained in the deposit with the ATCCDeposit Number shown for said cDNA clone in Table 1.

[0436] Also preferred is an isolated nucleic acid molecule, wherein saidnucleotide sequence encoding a polypeptide has been optimized forexpression of said polypeptide in a prokaryotic host.

[0437] Also preferred is an isolated nucleic acid molecule, wherein saidpolypeptide comprises an amino acid sequence selected from the groupconsisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is anyinteger as defined in Table 1; and a complete amino acid sequence of asecreted protein encoded by a human cDNA clone identified by a cDNAClone Identifier in Table 1 and contained in the deposit with the ATCCDeposit Number shown for said cDNA clone in Table 1.

[0438] Further preferred is a method of making a recombinant vectorcomprising inserting any of the above isolated nucleic acid moleculeinto a vector. Also preferred is the recombinant vector produced by thismethod. Also preferred is a method of making a recombinant host cellcomprising introducing the vector into a host cell, as well as therecombinant host cell produced by this method.

[0439] Also preferred is a method of making an isolated polypeptidecomprising culturing this recombinant host cell under conditions suchthat said polypeptide is expressed and recovering said polypeptide. Alsopreferred is this method of making an isolated polypeptide, wherein saidrecombinant host cell is a eukaryotic cell and said polypeptide is asecreted portion of a human secreted protein comprising an amino acidsequence selected from the group consisting of: an amino acid sequenceof SEQ ID NO:Y beginning with the residue at the position of the FirstAmino Acid of the Secreted Portion of SEQ ID NO:Y wherein Y is aninteger set forth in Table 1 and said position of the First Amino Acidof the Secreted Portion of SEQ ID NO:Y is defined in Table 1; and anamino acid sequence of a secreted portion of a protein encoded by ahuman cDNA clone identified by a cDNA Clone Identifier in Table 1 andcontained in the deposit with the ATCC Deposit Number shown for saidcDNA clone in Table 1. The isolated polypeptide produced by this methodis also preferred.

[0440] Also preferred is a method of treatment of an individual in needof an increased level of a secreted protein activity, which methodcomprises administering to such an individual a pharmaceuticalcomposition comprising an amount of an isolated polypeptide,polynucleotide, or antibody of the claimed invention effective toincrease the level of said protein activity in said individual.

[0441] Having generally described the invention, the same will be morereadily understood by reference to the following examples, which areprovided by way of illustration and are not intended as limiting.

EXAMPLES Example 1 Isolation of a Selected cDNA Clone From the DepositedSample

[0442] Each cDNA clone in a cited ATCC deposit is contained in a plasmidvector. Table 1 identifies the vectors used to construct the cDNAlibrary from which each clone was isolated. In many cases, the vectorused to construct the library is a phage vector from which a plasmid hasbeen excised. The table immediately below correlates the related plasmidfor each phage vector used in constructing the cDNA library. Forexample, where a particular clone is identified in Table 1 as beingisolated in the vector “Lambda Zap,” the corresponding deposited cloneis in “pBluescript.” Vector Used to Construct Library CorrespondingDeposited Plasmid Lambda Zap pBluescript (pBS) Uni-Zap XR pBluescript(pBS) Zap Express pBK lafmid BA plafmid BA pSport1 pSport1 pCMVSport 2.0pCMVSport 2.0 pCMVSport 3.0 pCMVSport 3.0 pCR ®2.1 pCR ®2.1

[0443] Vectors Lambda Zap (U.S. Pat. Nos. 5,128,256 and 5,286,636),Uni-Zap XR (U.S. Pat. Nos. 5,128,256 and 5,286,636), Zap Express (U.S.Pat. Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. etal., Nucleic Acids Res. 16:7583-7600 (1988); Alting-Mees, M. A. andShort, J. M., Nucleic Acids Res. 17:9494 (1989)) and pBK (Alting-Mees,M. A. et al., Strategies 5:58-61 (1992)) are commercially available fromStratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla,Calif., 92037. pBS contains an ampicillin resistance gene and pBKcontains a neomycin resistance gene. Both can be transformed into E.coli strain XL-1 Blue, also available from Stratagene. pBS comes in 4forms SK+, SK−, KS+ and KS. The S and K refers to the orientation of thepolylinker to the T7 and T3 primer sequences which flank the polylinkerregion (“S” is for SacI and “K” is for KpnI which are the first sites oneach respective end of the linker). “+” or “−” refer to the orientationof the f1 origin of replication (“ori” ), such that in one orientation,single stranded rescue initiated from the f1 ori generates sense strandDNA and in the other, antisense.

[0444] Vectors pSport1, pCMVSport 2.0 and pCMVSport 3.0, were obtainedfrom Life Technologies, Inc., P. O. Box 6009, Gaithersburg, Md. 20897.All Sport vectors contain an ampicillin resistance gene and may betransformed into E. coli strain DH10B, also available from LifeTechnologies. (See, for instance, Gruber, C. E., et al., Focus 15:59(1993).) Vector lafmid BA (Bento Soares, Columbia University, N.Y.)contains an ampicillin resistance gene and can be transformed into E.coli strain XL-1 Blue. Vector pCR®2.1, which is available fromInvitrogen, 1600 Faraday Avenue, Carlsbad, Calif. 92008, contains anampicillin resistance gene and may be transformed into E. coli strainDH10B, available from Life Technologies. (See, for instance, Clark, J.M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al.,Bio/Technology 9: (1991).) Preferably, a polynucleotide of the presentinvention does not comprise the phage vector sequences identified forthe particular clone in Table 1, as well as the corresponding plasmidvector sequences designated above.

[0445] The deposited material in the sample assigned the ATCC DepositNumber cited in Table 1 for any given cDNA clone also may contain one ormore additional plasmids, each comprising a cDNA clone different fromthat given clone. Thus, deposits sharing the same ATCC Deposit Numbercontain at least a plasmid for each cDNA clone identified in Table 1.Typically, each ATCC deposit sample cited in Table 1 comprises a mixtureof approximately equal amounts (by weight) of about 50 plasmid DNAs,each containing a different cDNA clone; but such a deposit sample mayinclude plasmids for more or less than 50 cDNA clones, up to about 500cDNA clones.

[0446] Two approaches can be used to isolate a particular clone from thedeposited sample of plasmid DNAs cited for that clone in Table 1. First,a plasmid is directly isolated by screening the clones using apolynucleotide probe corresponding to SEQ ID NO:X.

[0447] Particularly, a specific polynucleotide with 30-40 nucleotides issynthesized using an Applied Biosystems DNA synthesizer according to thesequence reported. The oligonucleotide is labeled, for instance, with³²P-γ-ATP using T4 polynucleotide kinase and purified according toroutine methods. (E.g., Maniatis et al., Molecular Cloning: A LaboratoryManual, Cold Spring Harbor Press, Cold Spring, N.Y. (1982).) The plasmidmixture is transformed into a suitable host, as indicated above (such asXL-1 Blue (Stratagene)) using techniques known to those of skill in theart, such as those provided by the vector supplier or in relatedpublications or patents cited above. The transformants are plated on1.5% agar plates (containing the appropriate selection agent, e.g.,ampicillin) to a density of about 150 transformants (colonies) perplate. These plates are screened using Nylon membranes according toroutine methods for bacterial colony screening (e.g., Sambrook et al.,Molecular Cloning: A Laboratory Manual, 2nd Edit., (1989), Cold SpringHarbor Laboratory Press, pages 1.93 to 1.104), or other techniques knownto those of skill in the art.

[0448] Alternatively, two primers of 17-20 nucleotides derived from bothends of the SEQ ID NO:X (i.e., within the region of SEQ ID NO:X boundedby the 5′ NT and the 3′ NT of the clone defined in Table 1) aresynthesized and used to amplify the desired cDNA using the depositedcDNA plasmid as a template. The polymerase chain reaction is carried outunder routine conditions, for instance, in 25 μl of reaction mixturewith 0.5 ug of the above cDNA template. A convenient reaction mixture is1.5-5 mM MgCl₂, 0.01% (w/v) gelatin, 20 μM each of dATP, dCTP, dGTP,dTTP, 25 pmol of each primer and 0.25 Unit of Taq polymerase. Thirtyfive cycles of PCR (denaturation at 94° C. for 1 min; annealing at 55°C. for 1 min; elongation at 72° C. for 1 min) are performed with aPerkin-Elmer Cetus automated thermal cycler. The amplified product isanalyzed by agarose gel electrophoresis and the DNA band with expectedmolecular weight is excised and purified. The PCR product is verified tobe the selected sequence by subcloning and sequencing the DNA product.

[0449] Several methods are available for the identification of the 5′ or3′ non-coding portions of a gene which may not be present in thedeposited clone. These methods include but are not limited to, filterprobing, clone enrichment using specific probes, and protocols similaror identical to 5′ and 3′ “RACE” protocols which are well known in theart. For instance, a method similar to 5′ RACE is available forgenerating the missing 5′ end of a desired full-length transcript.(Fromont-Racine et al., Nucleic Acids Res. 21(7):1683-1684 (1993).)

[0450] Briefly, a specific RNA oligonucleotide is ligated to the 5′ endsof a population of RNA presumably containing full-length gene RNAtranscripts. A primer set containing a primer specific to the ligatedRNA oligonucleotide and a primer specific to a known sequence of thegene of interest is used to PCR amplify the 5′ portion of the desiredfull-length gene. This amplified product may then be sequenced and usedto generate the full length gene.

[0451] This above method starts with total RNA isolated from the desiredsource, although poly-A+RNA can be used. The RNA preparation can then betreated with phosphatase if necessary to eliminate 5′ phosphate groupson degraded or damaged RNA which may interfere with the later RNA ligasestep. The phosphatase should then be inactivated and the RNA treatedwith tobacco acid pyrophosphatase in order to remove the cap structurepresent at the 5′ ends of messenger RNAs. This reaction leaves a 5′phosphate group at the 5′ end of the cap cleaved RNA which can then beligated to an RNA oligonucleotide using T4 RNA ligase.

[0452] This modified RNA preparation is used as a template for firststrand cDNA synthesis using a gene specific oligonucleotide. The firststrand synthesis reaction is used as a template for PCR amplification ofthe desired 5′ end using a primer specific to the ligated RNAoligonucleotide and a primer specific to the known sequence of the geneof interest. The resultant product is then sequenced and analyzed toconfirm that the 5′ end sequence belongs to the desired gene.

Example 2 Isolation of Genomic Clones Corresponding to a Polynucleotide

[0453] A human genomic P1 library (Genomic Systems, Inc.) is screened byPCR using primers selected for the cDNA sequence corresponding to SEQ IDNO:X., according to the method described in Example 1. (See also,Sambrook.)

Example 3 Tissue Distribution of Polypeptide

[0454] Tissue distribution of mRNA expression of polynucleotides of thepresent invention is determined using protocols for Northern blotanalysis, described by, among others, Sambrook et al. For example, acDNA probe produced by the method described in Example 1 is labeled withP³² using the rediprime™ DNA labeling system (Amersham Life Science),according to manufacturer's instructions. After labeling, the probe ispurified using CHROMA SPIN-100™ column (Clontech Laboratories, Inc.),according to manufacturer's protocol number PT1200-1. The purifiedlabeled probe is then used to examine various human tissues for mRNAexpression.

[0455] Multiple Tissue Northern (MTN) blots containing various humantissues (H) or human immune system tissues (IM) (Clontech) are examinedwith the labeled probe using ExpressHyb™ hybridization solution(Clontech) according to manufacturer's protocol number PT1190-1.Following hybridization and washing, the blots are mounted and exposedto film at −70° C. overnight, and the films developed according tostandard procedures.

Example 4 Chromosomal Mapping of the Polynucleotides

[0456] An oligonucleotide primer set is designed according to thesequence at the 5′ end of SEQ ID NO:X. This primer preferably spansabout 100 nucleotides. This primer set is then used in a polymerasechain reaction under the following set of conditions:30 seconds, 95° C.;1 minute, 56° C.; 1 minute, 70° C. This cycle is repeated 32 timesfollowed by one 5 minute cycle at 70° C. Human, mouse, and hamster DNAis used as template in addition to a somatic cell hybrid panelcontaining individual chromosomes or chromosome fragments (Bios, Inc).The reactions is analyzed on either 8% polyacrylamide gels or 3.5%agarose gels. Chromosome mapping is determined by the presence of anapproximately 100 bp PCR fragment in the particular somatic cell hybrid.

Example 5 Bacterial Expression or a Polypeptide

[0457] A polynucleotide encoding a polypeptide of the present inventionis amplified using PCR oligonucleotide primers corresponding to the 5′and 3′ ends of the DNA sequence, as outlined in Example 1, to synthesizeinsertion fragments. The primers used to amplify the cDNA insert shouldpreferably contain restriction sites, such as BamHI and XbaI, at the 5′end of the primers in order to clone the amplified product into theexpression vector. For example, BamHI and XbaI correspond to therestriction enzyme sites on the bacterial expression vector pQE-9.(Qiagen, Inc., Chatsworth, Calif.). This plasmid vector encodesantibiotic resistance (Amp^(r)), a bacterial origin of replication(ori), an IPTG-regulatable promoter/operator (P/O), a ribosome bindingsite (RBS), a 6-histidine tag (6-His), and restriction enzyme cloningsites.

[0458] The pQE-9 vector is digested with BamHI and XbaI and theamplified fragment is ligated into the pQE-9 vector maintaining thereading frame initiated at the bacterial RBS. The ligation mixture isthen used to transform the E. coli strain M15/rep4 (Qiagen, Inc.) whichcontains multiple copies of the plasmid pREP4, which expresses the lacIrepressor and also confers kanamycin resistance (Kan^(r)). Transformantsare identified by their ability to grow on LB plates andampicillin/kanamycin resistant colonies are selected. Plasmid DNA isisolated and confirmed by restriction analysis.

[0459] Clones containing the desired constructs are grown overnight(O/N) in liquid culture in LB media supplemented with both Amp (100ug/ml) and Kan (25 ug/ml). The O/N culture is used to inoculate a largeculture at a ratio of 1:100 to 1:250. The cells are grown to an opticaldensity 600 (O.D.⁶⁰⁰) of between 0.4 and 0.6. IPTG(Isopropyl-B-D-thiogalacto pyranoside) is then added to a finalconcentration of 1 mM. IPTG induces by inactivating the lacI repressor,clearing the P/O leading to increased gene expression.

[0460] Cells are grown for an extra 3 to 4 hours. Cells are thenharvested by centrifugation (20 mins at 6000 Xg). The cell pellet issolubilized in the chaotropic agent 6 Molar Guanidine HCl by stirringfor 3-4 hours at 4° C. The cell debris is removed by centrifugation, andthe supernatant containing the polypeptide is loaded onto anickel-nitrilo-tri-acetic acid (“Ni-NTA”) affinity resin column(available from QIAGEN, Inc., supra). Proteins with a 6×His tag bind tothe Ni-NTA resin with high affinity and can be purified in a simpleone-step procedure (for details see: The QIAexpressionist (1995) QIAGEN,Inc., supra).

[0461] Briefly, the supernatant is loaded onto the column in 6 Mguanidine-HCl, pH 8, the column is first washed with 10 volumes of 6 Mguanidine-HCl, pH 8, then washed with 10 volumes of 6 M guanidine-HCl pH6, and finally the polypeptide is eluted with 6 M guanidine-HCI, pH 5.

[0462] The purified protein is then renatured by dialyzing it againstphosphate-buffered saline (PBS) or 50 mM Na-acetate, pH 6 buffer plus200 mM NaCl. Alternatively, the protein can be successfully refoldedwhile immobilized on the Ni-NTA column. The recommended conditions areas follows: renature using a linear 6M-1M urea gradient in 500 mM NaCl,20% glycerol, 20 mM Tris/HCI pH 7.4, containing protease inhibitors. Therenaturation should be performed over a period of 1.5 hours or more.After renaturation the proteins are eluted by the addition of 250 mMimmidazole. Immidazole is removed by a final dialyzing step against PBSor 50 mM sodium acetate pH 6 buffer plus 200 mM NaCl. The purifiedprotein is stored at 4° C. or frozen at −80° C.

[0463] In addition to the above expression vector, the present inventionfurther includes an expression vector comprising phage operator andpromoter elements operatively linked to a polynucleotide of the presentinvention, called pHE4a. (ATCC Accession Number 209645, deposited onFeb. 25, 1998.) This vector contains: 1) a neomycinphosphotransferasegene as a selection marker, 2) an E. coli origin of replication, 3) a T5phage promoter sequence, 4) two lac operator sequences, 5) aShine-Delgarno sequence, and 6) the lactose operon repressor gene(lacIq). The origin of replication (oriC) is derived from pUC19 (LTI,Gaithersburg, Md.). The promoter sequence and operator sequences aremade synthetically.

[0464] DNA can be inserted into the pHEa by restricting the vector withNdeI and Xbal, BamHI, XhoI, or Asp718, running the restricted product ona gel, and isolating the larger fragment (the stuffer fragment should beabout 310 base pairs). The DNA insert is generated according to the PCRprotocol described in Example 1, using PCR primers having restrictionsites for NdeI (5′ primer) and XbaI, BamHI, XhoI, or Asp718 (3′ primer).The PCR insert is gel purified and restricted with compatible enzymes.The insert and vector are ligated according to standard protocols.

[0465] The engineered vector could easily be substituted in the aboveprotocol to express protein in a bacterial system.

Example 6 Purification of a Polypeptide from an Inclusion Body

[0466] The following alternative method can be used to purify apolypeptide expressed in E. coli when it is present in the form ofinclusion bodies. Unless otherwise specified, all of the following stepsare conducted at 4-10° C.

[0467] Upon completion of the production phase of the E. colifermentation, the cell culture is cooled to 4-10° C. and the cellsharvested by continuous centrifugation at 15,000 rpm (Heraeus Sepatech).On the basis of the expected yield of protein per unit weight of cellpaste and the amount of purified protein required, an appropriate amountof cell paste, by weight, is suspended in a buffer solution containing100 mM Tris, 50 mM EDTA, pH 7.4. The cells are dispersed to ahomogeneous suspension using a high shear mixer.

[0468] The cells are then lysed by passing the solution through amicrofluidizer (Microfuidics, Corp. or APV Gaulin, Inc.) twice at4000-6000 psi. The homogenate is then mixed with NaCl solution to afinal concentration of 0.5 M NaCI, followed by centrifugation at 7000×gfor 15 min. The resultant pellet is washed again using 0.5M NaCl, 100 mMTris, 50 mM EDTA, pH 7.4.

[0469] The resulting washed inclusion bodies are solubilized with 1.5 Mguanidine hydrochloride (GuHCI) for 2-4 hours. After 7000×gcentrifugation for 15 min., the pellet is discarded and the polypeptidecontaining supernatant is incubated at 4° C. overnight to allow furtherGuHCl extraction.

[0470] Following high speed centrifugation (30,000×g) to removeinsoluble particles, the GuHCI solubilized protein is refolded byquickly mixing the GuHCl extract with 20 volumes of buffer containing 50mM sodium, pH 4.5, 150 mM NaCl, 2 mM EDTA by vigorous stirring. Therefolded diluted protein solution is kept at 4° C. without mixing for 12hours prior to further purification steps.

[0471] To clarify the refolded polypeptide solution, a previouslyprepared tangential filtration unit equipped with 0.16 μm membranefilter with appropriate surface area (e.g., Filtron), equilibrated with40 mM sodium acetate, pH 6.0 is employed. The filtered sample is loadedonto a cation exchange resin (e.g., Poros HS-50, Perseptive Biosystems).The column is washed with 40 mM sodium acetate, pH 6.0 and eluted with250 mM, 500 mM, 1000 mM, and 1500 mM NaCl in the same buffer, in astepwise manner. The absorbance at 280 nm of the effluent iscontinuously monitored. Fractions are collected and further analyzed bySDS-PAGE.

[0472] Fractions containing the polypeptide are then pooled and mixedwith 4 volumes of water. The diluted sample is then loaded onto apreviously prepared set of tandem columns of strong anion (Poros HQ-50,Perseptive Biosystems) and weak anion (Poros CM-20, PerseptiveBiosystems) exchange resins. The columns are equilibrated with 40 mMsodium acetate, pH 6.0. Both columns are washed with 40 mM sodiumacetate, pH 6.0, 200 mM NaCl. The CM-20 column is then eluted using a 10column volume linear gradient ranging from 0.2 M NaCl, 50 mM sodiumacetate, pH 6.0 to 1.0 M NaCl, 50 mM sodium acetate, pH 6.5. Fractionsare collected under constant A₂₈₀ monitoring of the effluent. Fractionscontaining the polypeptide (determined, for instance, by 16% SDS-PAGE)are then pooled.

[0473] The resultant polypeptide should exhibit greater than 95% purityafter the above refolding and purification steps. No major contaminantbands should be observed from Commassie blue stained 16% SDS-PAGE gelwhen 5 μg of purified protein is loaded. The purified protein can alsobe tested for endotoxin/LPS contamination, and typically the LPS contentis less than 0.1 ng/ml according to LAL assays.

Example 7 Cloning and Expression of a Polypeptide in a BaculovirusExpression System

[0474] In this example, the plasmid shuttle vector pA2 is used to inserta polynucleotide into a baculovirus to express a polypeptide. Thisexpression vector contains the strong polyhedrin promoter of theAutographa californica nuclear polyhedrosis virus (AcMNPV) followed byconvenient restriction sites such as BamHI, Xba I and Asp718. Thepolyadenylation site of the simian virus 40 (“SV40”) is used forefficient polyadenylation. For easy selection of recombinant virus, theplasmid contains the beta-ogalactosidase gene from E. coli under controlof a weak Drosophila promoter in the same orientation, followed by thepolyadenylation signal of the polyhedrin gene. The inserted genes areflanked on both sides by viral sequences for cell-mediated homologousrecombination with wild-type viral DNA to generate a viable virus thatexpress the cloned polynucleotide.

[0475] Many other baculovirus vectors can be used in place of the vectorabove, such as pAc373, pVL941, and pAcIM1, as one skilled in the artwould readily appreciate, as long as the construct providesappropriately located signals for transcription, translation, secretionand the like, including a signal peptide and an in-frame AUG asrequired. Such vectors are described, for instance, in Luckow et al.,Virology 170:31-39 (1989).

[0476] Specifically, the cDNA sequence contained in the deposited clone,including the AUG initiation codon and the naturally associated leadersequence identified in Table 1, is amplified using the PCR protocoldescribed in Example 1 . If the naturally occurring signal sequence isused to produce the secreted protein, the pA2 vector does not need asecond signal peptide. Alternatively, the vector can be modified (pA2GP) to include a baculovirus leader sequence, using the standard methodsdescribed in Summers et al., “A Manual of Methods for BaculovirusVectors and Insect Cell Culture Procedures,” Texas AgriculturalExperimental Station Bulletin No. 1555 (1987).

[0477] The amplified fragment is isolated from a 1% agarose gel using acommercially available kit (“Geneclean,” BIO 101 Inc., La Jolla,Calif.). The fragment then is digested with appropriate restrictionenzymes and again purified on a 1% agarose gel.

[0478] The plasmid is digested with the corresponding restrictionenzymes and optionally, can be dephosphorylated using calf intestinalphosphatase, using routine procedures known in the art. The DNA is thenisolated from a 1% agarose gel using a commercially available kit(“Geneclean” BIO 101 Inc., La Jolla, Calif.).

[0479] The fragment and the dephosphorylated plasmid are ligatedtogether with T4 DNA ligase. E. coli HB101 or other suitable E. colihosts such as XL-1 Blue (Stratagene Cloning Systems, La Jolla, Calif.)cells are transformed with the ligation mixture and spread on cultureplates. Bacteria containing the plasmid are identified by digesting DNAfrom individual colonies and analyzing the digestion product by gelelectrophoresis. The sequence of the cloned fragment is confirmed by DNAsequencing.

[0480] Five μg of a plasmid containing the polynucleotide isco-transfected with 1.0 μg of a commercially available linearizedbaculovirus DNA (“BaculoGoId™ baculovirus DNA”, Pharmingen, San Diego,Calif.), using the lipofection method described by Felgner et al., Proc.Natl. Acad. Sci. USA 84:7413-7417 (1987). One μg of BaculoGold™ virusDNA and 5 μg of the plasmid are mixed in a sterile well of a microtiterplate containing 50 μl of serum-free Grace's medium (Life TechnologiesInc., Gaithersburg, Md.). Afterwards, 10 μl Lipofectin plus 90 μlGrace's medium are added, mixed and incubated for 15 minutes at roomtemperature. Then the transfection mixture is added drop-wise to Sf9insect cells (ATCC CRL 1711) seeded in a 35 mm tissue culture plate with1 ml Grace's medium without serum. The plate is then incubated for 5hours at 27° C. The transfection solution is then removed from the plateand 1 ml of Grace's insect medium supplemented with 10% fetal calf serumis added. Cultivation is then continued at 27° C. for four days.

[0481] After four days the supernatant is collected and a plaque assayis performed, as described by Summers and Smith, supra. An agarose gelwith “Blue Gal” (Life Technologies Inc., Gaithersburg) is used to alloweasy identification and isolation of gal-expressing clones, whichproduce blue-stained plaques. (A detailed description of a “plaqueassay” of this type can also be found in the user's guide for insectcell culture and baculovirology distributed by Life Technologies Inc.,Gaithersburg, page 9-10.) After appropriate incubation, blue stainedplaques are picked with the tip of a micropipettor (e.g., Eppendorf).The agar containing the recombinant viruses is then resuspended in amicrocentrifuge tube containing 200μl of Grace's medium and thesuspension containing the recombinant baculovirus is used to infect Sf9cells seeded in 35 mm dishes. Four days later the supernatants of theseculture dishes are harvested and then they are stored at 4° C.

[0482] To verify the expression of the polypeptide, Sf9 cells are grownin Grace's medium supplemented with 10% heat-inactivated FBS. The cellsare infected with the recombinant baculovirus containing thepolynucleotide at a multiplicity of infection (“MOI”) of about 2. Ifradiolabeled proteins are desired, 6 hours later the medium is removedand is replaced with SF900 II medium minus methionine and cysteine(available from Life Technologies Inc., Rockville, Md.). After 42 hours,5 μCi of ³⁵S-methionine and 5 μCi ³⁵S-cysteine (available from Amersham)are added. The cells are further incubated for 16 hours and then areharvested by centrifugation. The proteins in the supernatant as well asthe intracellular proteins are analyzed by SDS-PAGE followed byautoradiography (if radiolabeled).

[0483] Microsequencing of the amino acid sequence of the amino terminusof purified protein may be used to determine the amino terminal sequenceof the produced protein.

Example 8 Expression of a Polypeptide in Mammalian Cells

[0484] The polypeptide of the present invention can be expressed in amammalian cell. A typical mammalian expression vector contains apromoter element, which mediates the initiation of transcription ofmRNA, a protein coding sequence, and signals required for thetermination of transcription and polyadenylation of the transcript.Additional elements include enhancers, Kozak sequences and interveningsequences flanked by donor and acceptor sites for RNA splicing. Highlyefficient transcription is achieved with the early and late promotersfrom SV40, the long terminal repeats (LTRs) from Retroviruses, e.g.,RSV, HTLVI, HIVI and the early promoter of the cytomegalovirus (CMV).However, cellular elements can also be used (e.g., the human actinpromoter).

[0485] Suitable expression vectors for use in practicing the presentinvention include, for example, vectors such as pSVL and pMSG(Pharmacia, Uppsala, Sweden), pRSVcat (ATCC 37152), pSV2dhfr (ATCC37146), pBC12MI (ATCC 67109), pCMVSport 2.0, and pCMVSport 3.0.Mammalian host cells that could be used include, human Hela, 293, H9 andJurkat cells, mouse NIH3T3 and C127 cells, Cos 1, Cos 7 and CV1, quailQC1-3 cells, mouse L cells and Chinese hamster ovary (CHO) cells.

[0486] Alternatively, the polypeptide can be expressed in stable celllines containing the polynucleotide integrated into a chromosome. Theco-transfection with a selectable marker such as dhfr, gpt, neomycin,hygromycin allows the identification and isolation of the transfectedcells.

[0487] The transfected gene can also be amplified to express largeamounts of the encoded protein. The DHFR (dihydrofolate reductase)marker is useful in developing cell lines that carry several hundred oreven several thousand copies of the gene of interest. (See, e.g., Alt,F. W., et al., J. Biol. Chem. 253:1357-1370 (1978); Hamlin, J. L. andMa, C., Biochem. et Biophys. Acta, 1097:107-143 (1990); Page, M. J. andSydenham, M. A., Biotechnology 9:64-68 (1991).) Another useful selectionmarker is the enzyme glutamine synthase (GS) (Murphy et al., Biochem J.227:277-279 (1991); Bebbington et al., Bio/Technology 10:169-175 (1992).Using these markers, the mammalian cells are grown in selective mediumand the cells with the highest resistance are selected. These cell linescontain the amplified gene(s) integrated into a chromosome. Chinesehamster ovary (CHO) and NSO cells are often used for the production ofproteins.

[0488] Derivatives of the plasmid pSV2-dhfr (ATCC Accession No. 37146),the expression vectors pC4 (ATCC Accession No. 209646) and pC6 (ATCCAccession No.209647) contain the strong promoter (LTR) of the RousSarcoma Virus (Cullen et al., Molecular and Cellular Biology, 438-447(March, 1985)) plus a fragment of the CMV-enhancer (Boshart et al., Cell41:521-530 (1985).) Multiple cloning sites, e.g., with the restrictionenzyme cleavage sites BamHI, XbaI and Asp718, facilitate the cloning ofthe gene of interest. The vectors also contain the 3′ intron, thepolyadenylation and termination signal of the rat preproinsulin gene,and the mouse DHFR gene under control of the SV40 early promoter.

[0489] Specifically, the plasmid pC6, for example, is digested withappropriate restriction enzymes and then dephosphorylated using calfintestinal phosphates by procedures known in the art. The vector is thenisolated from a 1% agarose gel.

[0490] A polynucleotide of the present invention is amplified accordingto the protocol outlined in Example 1. If the naturally occurring signalsequence is used to produce the secreted protein, the vector does notneed a second signal peptide. Alternatively, if the naturally occurringsignal sequence is not used, the vector can be modified to include aheterologous signal sequence. (See, e.g., WO 96/34891.)

[0491] The amplified fragment is isolated from a 1% agarose gel using acommercially available kit (“Geneclean,” BIO 101 Inc., La Jolla,Calif.). The fragment then is digested with appropriate restrictionenzymes and again purified on a 1% agarose gel.

[0492] The amplified fragment is then digested with the same restrictionenzyme and purified on a 1% agarose gel. The isolated fragment and thedephosphorylated vector are then ligated with T4 DNA ligase. E. coliHB101 or XL-1 Blue cells are then transformed and bacteria areidentified that contain the fragment inserted into plasmid pC6 using,for instance, restriction enzyme analysis.

[0493] Chinese hamster ovary cells lacking an active DHFR gene is usedfor transfection. Five μg of the expression plasmid pC6 is cotransfectedwith 0.5 μg of the plasmid pSVneo using lipofectin (Feigner et al.,supra). The plasmid pSV2-neo contains a dominant selectable marker, theneo gene from Tn5 encoding an enzyme that confers resistance to a groupof antibiotics including G418. The cells are seeded in alpha minus MEMsupplemented with 1 mg/ml G418. After 2 days, the cells are trypsinizedand seeded in hybridoma cloning plates (Greiner, Germany) in alpha minusMEM supplemented with 10, 25, or 50 ng/ml of metothrexate plus 1 mg/mlG418. After about 10-14 days single clones are trypsinized and thenseeded in 6-well petri dishes or 10 ml flasks using differentconcentrations of methotrexate (50 nM, 100 nM, 200 nM, 400 nM, 800 nM).Clones growing at the highest concentrations of methotrexate are thentransferred to new 6-well plates containing even higher concentrationsof methotrexate (1 μM, 2 μM, 5 μM, 10 mM, 20 mM). The same procedure isrepeated until clones are obtained which grow at a concentration of100-200 μM. Expression of the desired gene product is analyzed, forinstance, by SDS-PAGE and Western blot or by reversed phase HPLCanalysis.

Example 9 Protein Fusions

[0494] The polypeptides of the present invention are preferably fused toother proteins. These fusion proteins can be used for a variety ofapplications. For example, fusion of the present polypeptides toHis-tag, HA-tag, protein A, IgG domains, and maltose binding proteinfacilitates purification. (See Example 5; see also EP A 394,827;Traunecker, et al., Nature 331:84-86 (1988).) Similarly, fusion toIgG-l, IgG-3, and albumin increases the halflife time in vivo. Nuclearlocalization signals fused to the polypeptides of the present inventioncan target the protein to a specific subcellular localization, whilecovalent heterodimer or homodimers can increase or decrease the activityof a fusion protein. Fusion proteins can also create chimeric moleculeshaving more than one function. Finally, fusion proteins can increasesolubility and/or stability of the fused protein compared to thenon-fused protein. All of the types of fusion proteins described abovecan be made by modifying the following protocol, which outlines thefusion of a polypeptide to an IgG molecule, or the protocol described inExample 5.

[0495] Briefly, the human Fc portion of the IgG molecule can be PCRamplified, using primers that span the 5′ and 3′ ends of the sequencedescribed below. These primers also should have convenient restrictionenzyme sites that will facilitate cloning into an expression vector,preferably a mammalian expression vector.

[0496] For example, if pC4 (Accession No. 209646) is used, the human Fcportion can be ligated into the BamHI cloning site. Note that the 3′BamHI site should be destroyed. Next, the vector containing the human Fcportion is re-restricted with BamHI, linearizing the vector, and apolynucleotide of the present invention, isolated by the PCR protocoldescribed in Example 1, is ligated into this BamHI site. Note that thepolynucleotide is cloned without a stop codon, otherwise a fusionprotein will not be produced.

[0497] If the naturally occurring signal sequence is used to produce thesecreted protein, pC4 does not need a second signal peptide.Alternatively, if the naturally occurring signal sequence is not used,the vector can be modified to include a heterologous signal sequence.(See, e.g., WO 96/34891.)

[0498] Human IgG Fc region: GGGATCCGGAGCCCAAATCTTCTGACAAAACTCACAC (SEQID NO:1) ATGCCCACCGTGCCCAGCACCTGAATTCGAGGGTGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACTCCTGAGGTCACATGCGTGGTGGTGGACGTAAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAACCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCAAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAA ATGAGTGCGACGGCCGCGACTCTAGAGGAT

Example 10 Production of an Antibody from a Polypeptide

[0499] The antibodies of the present invention can be prepared by avariety of methods. (See, Current Protocols, Chapter 2.) For example,cells expressing a polypeptide of the present invention is administeredto an animal to induce the production of sera containing polyclonalantibodies. In a preferred method, a preparation of the secreted proteinis prepared and purified to render it substantially free of naturalcontaminants. Such a preparation is then introduced into an animal inorder to produce polyclonal antisera of greater specific activity.

[0500] In the most preferred method, the antibodies of the presentinvention are monoclonal antibodies (or protein binding fragmentsthereof). Such monoclonal antibodies can be prepared using hybridomatechnology. (Köhler et al., Nature 256:495 (1975); Köhler et al., Eur.J. Immunol. 6:511 (1976); Köohler et al., Eur. J. Immunol. 6:292 (1976);Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas,Elsevier, N.Y., pp. 563-681 (1981).) In general, such procedures involveimmunizing an animal (preferably a mouse) with polypeptide or, morepreferably, with a secreted polypeptide-expressing cell. Such cells maybe cultured in any suitable tissue culture medium; however, it ispreferable to culture cells in Earle's modified Eagle's mediumsupplemented with 10% fetal bovine serum (inactivated at about 56° C.),and supplemented with about 10 g/l of nonessential amino acids, about1,000 U/ml of penicillin, and about 100 μg/ml of streptomycin.

[0501] The splenocytes of such mice are extracted and fused with asuitable myeloma cell line. Any suitable myeloma cell line may beemployed in accordance with the present invention; however, it ispreferable to employ the parent myeloma cell line (SP20), available fromthe ATCC. After fusion, the resulting hybridoma cells are selectivelymaintained in HAT medium, and then cloned by limiting dilution asdescribed by Wands et al. (Gastroenterology 80:225-232 (1981).) Thehybridoma cells obtained through such a selection are then assayed toidentify clones which secrete antibodies capable of binding thepolypeptide.

[0502] Alternatively, additional antibodies capable of binding to thepolypeptide can be produced in a two-step procedure using anti-idiotypicantibodies. Such a method makes use of the fact that antibodies arethemselves antigens, and therefore, it is possible to obtain an antibodywhich binds to a second antibody. In accordance with this method,protein specific antibodies are used to immunize an animal, preferably amouse. The splenocytes of such an animal are then used to producehybridoma cells, and the hybridoma cells are screened to identify cloneswhich produce an antibody whose ability to bind to the protein-specificantibody can be blocked by the polypeptide. Such antibodies compriseanti-idiotypic antibodies to the protein-specific antibody and can beused to immunize an animal to induce formation of furtherprotein-specific antibodies.

[0503] It will be appreciated that Fab and F(ab′)2 and other fragmentsof the antibodies of the present invention may be used according to themethods disclosed herein. Such fragments are typically produced byproteolytic cleavage, using enzymes such as papain (to produce Fabfragments) or pepsin (to produce F(ab′)2 fragments). Alternatively,secreted protein-binding fragments can be produced through theapplication of recombinant DNA technology or through syntheticchemistry.

[0504] For in vivo use of antibodies in humans, it may be preferable touse “humanized” chimeric monoclonal antibodies. Such antibodies can beproduced using genetic constructs derived from hybridoma cells producingthe monoclonal antibodies described above. Methods for producingchimeric antibodies are known in the art. (See, for review, Morrison,Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Cabillyet al., U.S. Pat. No. 4,816,567; Taniguchi et al., EP 171496; Morrisonet al., EP 173494; Neuberger et al., WO 8601533; Robinson et al., WO8702671; Boulianne et al., Nature 312:643 (1984); Neuberger et al.,Nature 314:268 (1985).)

Example 11 Production Of Secreted Protein For High-Throughput ScreeningAssays

[0505] The following protocol produces a supernatant containing apolypeptide to be tested. This supernatant can then be used in theScreening Assays described in Examples 13-20.

[0506] First, dilute Poly-D-Lysine (644 587 Boehringer-Mannheim) stocksolution (1 mg/ml in PBS) 1:20 in PBS (w/o calcium or magnesium 17-516FBiowhittaker) for a working solution of 50 ug/ml. Add 200 ul of thissolution to each well (24 well plates) and incubate at RT for 20minutes. Be sure to distribute the solution over each well (note: a12-channel pipetter may be used with tips on every other channel).Aspirate off the Poly-D-Lysine solution and rinse with 1 ml PBS(Phosphate Buffered Saline). The PBS should remain in the well untiljust prior to plating the cells and plates may be poly-lysine coated inadvance for up to two weeks.

[0507] Plate 293T cells (do not carry cells past P+20) at 2×10⁵cells/well in 0.5 ml DMEM(Dulbecco's Modified Eagle Medium)(with 4.5 G/Lglucose and L-glutamine (12-604F Biowhittaker))/10% heat inactivatedFBS(14-503F Biowhittaker)/1×Penstrep(17-602E Biowhittaker). Let thecells grow overnight.

[0508] The next day, mix together in a sterile solution basin: 300 ulLipofectamine (18324-012 Gibco/BRL) and 5 ml Optimem I (31985070Gibco/BRL)/96-well plate. With a small volume multi-channel pipetter,aliquot approximately 2 ug of an expression vector containing apolynucleotide insert, produced by the methods described in Examples 8or 9, into an appropriately labeled 96-well round bottom plate. With amulti-channel pipetter, add 50 ul of the Lipofectamine/Optimem I mixtureto each well. Pipette up and down gently to mix. Incubate at RT 15-45minutes. After about 20 minutes, use a multi-channel pipetter to add 150ul Optimem I to each well. As a control, one plate of vector DNA lackingan insert should be transfected with each set of transfections.

[0509] Preferably, the transfection should be performed by tag-teamingthe following tasks. By tag-teaming, hands on time is cut in half, andthe cells do not spend too much time on PBS. First, person A aspiratesoff the media from four 24-well plates of cells, and then person Brinses each well with 0.5-1 ml PBS. Person A then aspirates off PBSrinse, and person B, using a12-channel pipetter with tips on every otherchannel, adds the 200 ul of DNA/Lipofectamine/Optimem I complex to theodd wells first, then to the even wells, to each row on the 24-wellplates. Incubate at 37° C. for 6 hours.

[0510] While cells are incubating, prepare appropriate media, either1%BSA in DMEM with 1×penstrep, or CHO-5 media (116.6 mg/L of CaCl2(anhyd); 0.00130 mg/L CuSO₄-5H₂O; 0.050 mg/L of Fe(NO₃)₃-9H₂O; 0.417mg/L of FeSO₄-7H₂O; 311.80 mg/L of Kcl; 28.64 mg/L of MgCl₂; 48.84 mg/Lof MgSO₄; 6995.50 mg/L of NaCl; 2400.0 mg/L of NaHCO₃; 62.50 mg/L ofNaH₂PO₄-H₂O ; 71.02 mg/L of Na₂HPO4; 0.4320 mg/L of ZnSO₄-7H₂O; 0.002mg/L of Arachidonic Acid; 1.022 mg/L of Cholesterol; 0.070 mg/L ofDL-alpha-Tocopherol-Acetate; 0.0520 mg/L of Linoleic Acid; 0.010 mg/L ofLinolenic Acid; 0.010 mg/L of Myristic Acid; 0.010 mg/L of Oleic Acid;0.010 mg/L of Palmitric Acid; 0.010 mg/L of Palmitic Acid; 100 mg/L ofPluronic F-68; 0.010 mg/L of Stearic Acid; 2.20 mg/L of Tween 80; 4551mg/L of D-Glucose; 130.85 mg/ml of L- Alanine; 147.50 mg/ml ofL-Arginine-HCL; 7.50 mg/ml of L-Asparagine-H₂O; 6.65 mg/ml of L-AsparticAcid; 29.56 mg/ml of L-Cystine-2HCL-H₂O; 31.29 mg/ml of L-Cystine-2HCL;7.35 mg/ml of L-Glutamic Acid; 365.0 mg/ml of L-Glutamine; 18.75 mg/mlof Glycine; 52.48 mg/ml of L-Histidine-HCL-H₂O; 106.97 mg/ml ofL-Isoleucine; 111.45 mg/ml of L-Leucine; 163.75 mg/ml of L-Lysine HCL;32.34 mg/ml of L-Methionine; 68.48 mg/ml of L-Phenylalainine; 40.0 mg/mlof L-Proline; 26.25 mg/ml of L-Serine; 101.05 mg/ml of L-Threonine;19.22 mg/ml of L-Tryptophan; 91.79 mg/ml of L-Tryrosine-2Na-2H₂O; 99.65mg/ml of L-Valine; 0.0035 mg/L of Biotin; 3.24 mg/L of D-CaPantothenate; 11.78 mg/L of Choline Chloride; 4.65 mg/L of Folic Acid;15.60 mg/L of i-Inositol; 3.02 mg/L of Niacinamide; 3.00 mg/L ofPyridoxal HCL; 0.031 mg/L of Pyridoxine HCL; 0.319 mg/L of Riboflavin;3.17 mg/L of Thiamine HCL; 0.365 mg/L of Thymidine; and 0.680 mg/L ofVitamin B₁₂; 25 mM of HEPES Buffer; 2.39 mg/L of Na Hypoxanthine; 0.105mg/L of Lipoic Acid; 0.081 mg/L of Sodium Putrescine-2HCL; 55.0 mg/L ofSodium Pyruvate; 0.0067 mg/L of Sodium Selenite; 20 uM of Ethanolamine;0.122 mg/L of Ferric Citrate; 41.70 mg/L of Methyl-B-Cyclodextrincomplexed with Linoleic Acid; 33.33 mg/L of Methyl-B-Cyclodextrincomplexed with Oleic Acid; and 10 mg/L of Methyl-B-Cyclodextrincomplexed with Retinal) with 2 mm glutamine and 1×penstrep. (BSA(81-068-3 Bayer) 100 gm dissolved in 1L DMEM for a 10% BSA stocksolution). Filter the media and collect 50 ul for endotoxin assay n l15ml polystyrene conical.

[0511] The transfection reaction is terminated, preferably bytag-teaming, at the end of the incubation period. Person A aspirates offthe transfection media, while person B adds 1.5 ml appropriate media toeach well. Incubate at 37° C. for 45 or 72 hours depending on the mediaused: 1% BSA for 45 hours or CHO-5 for 72 hours.

[0512] On day four, using a 300 ul multichannel pipetter, aliquot 600 ulin one 1 ml deep well plate and the remaining supernatant into a 2 mldeep well. The supernatants from each well can then be used in theassays described in Examples 13-20.

[0513] It is specifically understood that when activity is obtained inany of the assays described below using a supernatant, the activityoriginates from either the polypeptide directly (e.g., as a secretedprotein) or by the polypeptide inducing expression of other proteins,which are then secreted into the supernatant. Thus, the inventionfurther provides a method of identifying the protein in the supernatantcharacterized by an activity in a particular assay.

Example 12 Construction of GAS Reporter Construct

[0514] One signal transduction pathway involved in the differentiationand proliferation of cells is called the Jaks-STATs pathway. Activatedproteins in the Jaks-STATs pathway bind to gamma activation site “GAS”elements or interferon-sensitive responsive element (“ISRE”), located inthe promoter of many genes. The binding of a protein to these elementsalter the expression of the associated gene.

[0515] GAS and ISRE elements are recognized by a class of transcriptionfactors called Signal Transducers and Activators of Transcription, or“STATs.” There are six members of the STATs family. Stat1 and Stat3 arepresent in many cell types, as is Stat2 (as response to IFN-alpha iswidespread). Stat4 is more restricted and is not in many cell typesthough it has been found in T helper class I, cells after treatment withIL-12. Stat5 was originally called mammary growth factor, but has beenfound at higher concentrations in other cells including myeloid cells.It can be activated in tissue culture cells by many cytokines.

[0516] The STATs are activated to transiocate from the cytoplasm to thenucleus upon tyrosine phosphorylation by a set of kinases known as theJanus Kináse (“Jaks”) family. Jaks represent a distinct family ofsoluble tyrosine kinases and include Tyk2, Jak1, Jak2, and Jak3. Thesekinases display significant sequence similarity and are generallycatalytically inactive in resting cells.

[0517] The Jaks are activated by a wide range of receptors summarized inthe Table below. (Adapted from review by Schidler and Darnell, Ann. Rev.Biochem. 64:621-51 (1995).) A cytokine receptor family, capable ofactivating Jaks, is divided into two groups: (a) Class 1 includesreceptors for IL-2, IL-3, IL-4, IL-6, IL-7, IL-9, IL-11, IL-12, IL-15,Epo, PRL, GH, G-CSF, GM-CSF, LIF, CNTF, and thrombopoietin; and (b)Class 2 includes IFN-a, IFN-g, and IL-10. The Class 1 receptors share aconserved cysteine motif (a set of four conserved cysteines and onetryptophan) and a WSXWS motif (a membrane proxial region encodingTrp-Ser-Xxx-Trp-Ser (SEQ ID NO:2))

[0518] Thus, on binding of a ligand to a receptor, Jaks are activated,which in turn activate STATs, which then translocate and bind to GASelements. This entire process is encompassed in the Jaks-STATs signaltransduction pathway.

[0519] Therefore, activation of the Jaks-STATs pathway, reflected by thebinding of the GAS or the ISRE element, can be used to indicate proteinsinvolved in the proliferation and differentiation of cells. For example,growth factors and cytokines are known to activate the Jaks-STATspathway. (See Table below.) Thus, by using GAS elements linked toreporter molecules, activators of the Jaks-STATs pathway can beidentified. JAKs Ligand tyk2 Jak1 Jak2 Jak3 STATS GAS (elements) or ISREIFN family IFN-a/B + + − − 1,2,3 ISRE IFN-g + + − 1 GAS (IRF1 > Lys6 >IFP) Il-10 + ? ? − 1,3 gp130 family IL-6 (Pleiotropic) + + + ? 1,3 GAS(IRF1 > Lys6 > IFP) Il-11 (Pleiotropic) ? + ? ? 1,3 OnM (Pleiotropic)? + + ? 1,3 LIF (Pleiotropic) ? + + ? 1,3 CNTF (Pleiotropic) −/+ + + ?1,3 G-CSF (Pleiotropic) ? + ? ? 1,3 IL-12 (Pleiotropic) + − + + 1,3 g-Cfamily IL-2 (lymphocytes) − + − + 1,3,5 GAS IL-4 (lymph/myeloid) − + − +6 GAS (IRF1 = IFP >> Ly6)(IgH) IL-7 (lymphocytes) − + − + 5 GAS IL-9(lymphocytes) − + − + 5 GAS IL-13 (lymphocyte) − + ? ? 6 GAS IL-15 ? +? + 5 GAS gp140 family IL-3 (myeloid) − − + − 5 GAS (IRF1 > IFP >> Ly6)IL-5 (myeloid) − − + − 5 GAS GM-CSF (myeloid) − − + − 5 GAS Growthhormone family GH ? − + − 5 PRL ? +/− + − 1,3,5 EPO ? − + − 5 GAS(B-CAS > IRF1 = IFP >> Ly6) Receptor Tyrosine Kinases EGF ? + + − 1,3GAS (IRF1) PDGF ? + + − 1,3 CSF-1 ? + + − 1,3 GAS (not IRF1)

[0520] To construct a synthetic GAS containing promoter element, whichis used in the Biological Assays described in Examples 13-14, a PCRbased strategy is employed to generate a GAS-SV40 promoter sequence. The5′ primer contains four tandem copies of the GAS binding site found inthe IRF1 promoter and previously demonstrated to bind STATs uponinduction with a range of cytokines (Rothman et al., Immunity 1:457-468(1994).), although other GAS or ISRE elements can be used instead. The5′ primer also contains 18 bp of sequence complementary to the SV40early promoter sequence and is flanked with an XhoI site. The sequenceof the 5′ primer is:

[0521] 5′:GCGCCTCGAGATTTCCCCGAAATCTAGATTTCCCCGAAATGATTTCCCCGAAATGATTTCCCCGAAATATCTGCCATCTCAATTAG:3′ (SEQ ID NO:3)

[0522] The downstream primer is complementary to the SV40 promoter andis flanked with a Hind III site: 5′:GCGGCAAGCTTTTTGCAAAGCCTAGGC:3′ (SEQID NO:4)

[0523] PCR amplification is performed using the SV40 promoter templatepresent in the B-gal:promoter plasmid obtained from Clontech. Theresulting PCR fragment is digested with XhoI/Hind III and subcloned intoBLSK2-. (Stratagene.) Sequencing with forward and reverse primersconfirms that the insert contains the following sequence:5′:CTCGAGATTTCCCCGAAATCTAGATTTCCCCGAA (SEQ ID NO:5)ATGATTTCCCCGAAATGATTTCCCCGAAATATCTGCCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGC TTTTGCAAA+E,UNC AAGCTTT:3′

[0524] With this GAS promoter element linked to the SV40 promoter, aGAS:SEAP2 reporter construct is next engineered. Here, the reportermolecule is a secreted alkaline phosphatase, or “SEAP.” Clearly,however, any reporter molecule can be instead of SEAP, in this or in anyof the other Examples. Well known reporter molecules that can be usedinstead of SEAP include chloramphenicol acetyltransferase (CAT),luciferase, alkaline phosphatase, B-galactosidase, green fluorescentprotein (GFP), or any protein detectable by an antibody.

[0525] The above sequence confirmed synthetic GAS-SV40 promoter elementis subcloned into the pSEAP-Promoter vector obtained from Clontech usingHindIII and XhoI, effectively replacing the SV40 promoter with theamplified GAS:SV40 promoter element, to create the GAS-SEAP vector.However, this vector does not contain a neomycin resistance gene, andtherefore, is not preferred for mammalian expression systems.

[0526] Thus, in order to generate mammalian stable cell lines expressingthe GAS-SEAP reporter, the GAS-SEAP cassette is removed from theGAS-SEAP vector using SalI and NotI, and inserted into a backbone vectorcontaining the neomycin resistance gene, such as pGFP-1 (Clontech),using these restriction sites in the multiple cloning site, to createthe GAS-SEAP/Neo vector. Once this vector is transfected into mammaliancells, this vector can then be used as a reporter molecule for GASbinding as described in Examples 13-14.

[0527] Other constructs can be made using the above description andreplacing GAS with a different promoter sequence. For example,construction of reporter molecules containing NFK-B and EGR promotersequences are described in Examples 15 and 16. However, many otherpromoters can be substituted using the protocols described in theseExamples. For instance, SRE, IL-2, NFAT, or Osteocalcin promoters can besubstituted, alone or in combination (e.g., GAS/NF-KB/EGR, GAS/NF-KB,Il-2/NFAT, or NF-KB/GAS). Similarly, other cell lines can be used totest reporter construct activity, such as HELA (epithelial), HUVEC(endothelial), Reh (B-cell), Saos-2 (osteoblast), HUVAC (aortic), orCardiomyocyte.

Example 13 High-Throughput Screening Assay for T-cell Activity

[0528] The following protocol is used to assess T-cell activity byidentifying factors, such as growth factors and cytokines, that mayproliferate or differentiate T-cells. T-cell activity is assessed usingthe GAS/SEAP/Neo construct produced in Example 12. Thus, factors thatincrease SEAP activity indicate the ability to activate the Jaks-STATSsignal transduction pathway. The T-cell used in this assay is JurkatT-cells (ATCC Accession No. TIB-152), although Molt-3 cells (ATCCAccession No. CRL-1552) and Molt-4 cells (ATCC Accession No. CRL-1582)cells can also be used.

[0529] Jurkat T-cells are lymphoblastic CD4+Th1 helper cells. In orderto generate stable cell lines, approximately 2 million Jurkat cells aretransfected with the GAS-SEAP/neo vector using DMRIE-C (LifeTechnologies)(transfection procedure described below). The transfectedcells are seeded to a density of approximately 20,000 cells per well andtransfectants resistant to 1 mg/ml genticin selected. Resistant coloniesare expanded and then tested for their response to increasingconcentrations of interferon gamma. The dose response of a selectedclone is demonstrated.

[0530] Specifically, the following protocol will yield sufficient cellsfor 75 wells containing 200 ul of cells. Thus, it is either scaled up,or performed in multiple to generate sufficient cells for multiple 96well plates. Jurkat cells are maintained in RPMI+10% serum with 1%Pen-Strep. Combine 2.5 mls of OPTI-MEM (Life Technologies) with 10 ug ofplasmid DNA in a T25 flask. Add 2.5 ml OPTI-MEM containing 50 ul ofDMRIE-C and incubate at room temperature for 15-45 mins.

[0531] During the incubation period, count cell concentration, spin downthe required number of cells (10⁷ per transfection), and resuspend inOPTI-MEM to a final concentration of 10⁷ cells/mi. Then add 1 ml of1×10⁷ cells in OPTI-MEM to T25 flask and incubate at 37° C. for 6 hrs.After the incubation, add 10 ml of RPMI+15% serum.

[0532] The Jurkat:GAS-SEAP stable reporter lines are maintained inRPMI+10% serum, 1 mg/ml Genticin, and 1% Pen-Strep. These cells aretreated with supernatants containing a polypeptide as produced by theprotocol described in Example 11.

[0533] On the day of treatment with the supernatant, the cells should bewashed and resuspended in fresh RPMI+10% serum to a density of 500,000cells per ml. The exact number of cells required will depend on thenumber of supernatants being screened. For one 96 well plate,approximately 10 million cells (for 10 plates, 100 million cells) arerequired.

[0534] Transfer the cells to a triangular reservoir boat, in order todispense the cells into a 96 well dish, using a 12 channel pipette.Using a 12 channel pipette, transfer 200 ul of cells into each well(therefore adding 100, 000 cells per well).

[0535] After all the plates have been seeded, 50 ul of the supernatantsare transferred directly from the 96 well plate containing thesupernatants into each well using a 12 channel pipette. In addition, adose of exogenous interferon gamma (0.1, 1.0, 10 ng) is added to wellsH9, H10, and H11 to serve as additional positive controls for the assay.

[0536] The 96 well dishes containing Jurkat cells treated withsupernatants are placed in an incubator for 48 hrs (note: this time isvariable between 48-72 hrs). 35 ul samples from each well are thentransferred to an opaque 96 well plate using a 12 channel pipette. Theopaque plates should be covered (using sellophene covers) and storedat—20° C. until SEAP assays are performed according to Example 17. Theplates containing the remaining treated cells are placed at 4° C. andserve as a source of material for repeating the assay on a specific wellif desired.

[0537] As a positive control, 100 Unit/ml interferon gamma can be usedwhich is known to activate Jurkat T cells. Over 30 fold induction istypically observed in the positive control wells.

Example 14 High-Throughput Screening Assay Identifying Myeloid Activity

[0538] The following protocol is used to assess myeloid activity byidentifying factors, such as growth factors and cytokines, that mayproliferate or differentiate myeloid cells. Myeloid cell activity isassessed using the GAS/SEAP/Neo construct produced in Example 12. Thus,factors that increase SEAP activity indicate the ability to activate theJaks-STATS signal transduction pathway. The myeloid cell used in thisassay is U937, a pre-monocyte cell line, although TF-1, HL60, or KG1 canbe used.

[0539] To transiently transfect U937 cells with the GAS/SEAP/Neoconstruct produced in Example 12, a DEAE-Dextran method (Kharbanda et.al., 1994, Cell Growth & Differentiation, 5:259-265) is used. First,harvest 2×10e⁷ U937 cells and wash with PBS. The U937 cells are usuallygrown in RPMI 1640 medium containing 10% heat-inactivated fetal bovineserum (FBS) supplemented with 100 units/ml penicillin and 100 mg/mlstreptomycin.

[0540] Next, suspend the cells in 1 ml of 20 mM Tris-HCl (pH 7.4) buffercontaining 0.5 mg/ml DEAE-Dextran, 8 ug GAS-SEAP2 plasmid DNA, 140 mMNaCl, 5 mM KCl, 375 uM Na₂HPO₄. 7H₂O, 1 mM MgCl₂, and 675 uM CaCl₂.Incubate at 37° C. for 45 min.

[0541] Wash the cells with RPMI 1640 medium containing 10% FBS and thenresuspend in 10 ml complete medium and incubate at 37° C. for 36 hr.

[0542] The GAS-SEAP/U937 stable cells are obtained by growing the cellsin 400 ug/ml G418. The G418-free medium is used for routine growth butevery one to two months, the cells should be re-grown in 400 ug/ml G418for couple of passages.

[0543] These cells are tested by harvesting 1×10⁸ cells (this is enoughfor ten 96-well plates assay) and wash with PBS. Suspend the cells in200 ml above described growth medium, with a final density of 5×10⁵cells/ml. Plate 200 ul cells per well in the 96-well plate (or 1×10⁵cells/well).

[0544] Add 50 ul of the supernatant prepared by the protocol describedin Example 11. Incubate at 37° C. for 48 to 72 hr. As a positivecontrol, 100 Unit/ml interferon gamma can be used which is known toactivate U937 cells. Over 30 fold induction is typically observed in thepositive control wells. SEAP assay the supernatant according to theprotocol described in Example 17.

Example 15 High-Throughput Screening Assay Identifying NeuronalActivity.

[0545] When cells undergo differentiation and proliferation, a group ofgenes are activated through many different signal transduction pathways.One of these genes, EGR1 (early growth response gene 1), is induced invarious tissues and cell types upon activation. The promoter of EGR1 isresponsible for such induction. Using the EGR1 promoter linked toreporter molecules, activation of cells can be assessed.

[0546] Particularly, the following protocol is used to assess neuronalactivity in PC12 cell lines. PC12 cells (rat phenochromocytoma cells)are known to proliferate and/or differentiate by activation with anumber of mitogens, such as TPA (tetradecanoyl phorbol acetate), NGF(nerve growth factor), and EGF (epidermal growth factor). The EGR1 geneexpression is activated during this treatment. Thus, by stablytransfecting PC12 cells with a construct containing an EGR promoterlinked to SEAP reporter, activation of PC12 cells can be assessed.

[0547] The EGR/SEAP reporter construct can be assembled by the followingprotocol. The EGR-1 promoter sequence (−633 to +1)(Sakamoto K et al.,Oncogene 6:867-871 (1991)) can be PCR amplified from human genomic DNAusing the following primers: 5′GCGCTCGAGGGATGACAGCGATAGAACCCCGG-3′ (SEQID NO:6) 5′GCGAAGCTTCGCGACTCCCCGGATCCGCCTC-3′ (SEQ ID NO:7)

[0548] Using the GAS:SEAP/Neo vector produced in Example 12, EGR1amplified product can then be inserted into this vector. Linearize theGAS:SEAP/Neo vector using restriction enzymes XhoI/HindIII, removing theGAS/SV40 stuffer. Restrict the EGR1 amplified product with these sameenzymes. Ligate the vector and the EGR1 promoter.

[0549] To prepare 96 well-plates for cell culture, two mls of a coatingsolution (1:30 dilution of collagen type I (Upstate Biotech Inc.Cat#08-115) in 30% ethanol (filter sterilized)) is added per one 10 cmplate or 50 ml per well of the 96-well plate, and allowed to air dry for2 hr.

[0550] PC12 cells are routinely grown in RPMI-1640 medium (BioWhittaker) containing 10% horse serum (JRH BIOSCIENCES, Cat. #12449-78P), 5% heat-inactivated fetal bovine serum (FBS) supplementedwith 100 units/ml penicillin and 100 ug/ml streptomycin on a precoated10 cm tissue culture dish. One to four split is done every three to fourdays. Cells are removed from the plates by scraping and resuspended withpipetting up and down for more than 15 times.

[0551] Transfect the EGR/SEAP/Neo construct into PC12 using theLipofectamine protocol described in Example 11. EGR-SEAP/PC12 stablecells are obtained by growing the cells in 300 ug/ml G418. The G418-freemedium is used for routine growth but every one to two months, the cellsshould be re-grown in 300 ug/ml G418 for couple of passages.

[0552] To assay for neuronal activity, a 10 cm plate with cells around70 to 80% confluent is screened by removing the old medium. Wash thecells once with PBS (Phosphate buffered saline). Then starve the cellsin low serum medium (RPMI-1640 containing 1% horse serum and 0.5% FBSwith antibiotics) overnight.

[0553] The next morning, remove the medium and wash the cells with PBS.Scrape off the cells from the plate, suspend the cells well in 2 ml lowserum medium. Count the cell number and add more low serum medium toreach final cell density as 5×10⁵ cells/ml.

[0554] Add 200 ul of the cell suspension to each well of 96-well plate(equivalent to 1×10⁵ cells/well). Add 50 ul supernatant produced byExample 11, 37° C. for 48 to 72 hr. As a positive control, a growthfactor known to activate PC12 cells through EGR can be used, such as 50ng/ul of Neuronal Growth Factor (NGF). Over fifty-fold induction of SEAPis typically seen in the positive control wells. SEAP assay thesupernatant according to Example 17.

Example 16 High-Throughput Screening Assay for T-cell Activity

[0555] NF-κB (Nuclear Factor κB) is a transcription factor activated bya wide variety of agents including the inflammatory cytokines IL-1 andTNF, CD30 and CD40, lymphotoxin-alpha and lymphotoxin-beta, by exposureto LPS or thrombin, and by expression of certain viral gene products. Asa transcription factor, NF-κB regulates the expression of genes involvedin immune cell activation, control of apoptosis (NF-κB appears to shieldcells from apoptosis), B and T-cell development, anti-viral andantimicrobial responses, and multiple stress responses.

[0556] In non-stimulated conditions, NF-κB is retained in the cytoplasmwith I-κB (Inhibitor κB). However, upon stimulation, I-κB isphosphorylated and degraded, causing NF-κB to shuttle to the nucleus,thereby activating transcription of target genes. Target genes activatedby NF-κB include IL-2, IL-6, GM-CSF, ICAM-1 and class 1 MHC.

[0557] Due to its central role and ability to respond to a range ofstimuli, reporter constructs utilizing the NF-κB promoter element areused to screen the supernatants produced in Example 11. Activators orinhibitors of NF-kB would be useful in treating diseases. For example,inhibitors of NF-κB could be used to treat those diseases related to theacute or chronic activation of NF-kB, such as rheumatoid arthritis.

[0558] To construct a vector containing the NF-κB promoter element, aPCR based strategy is employed. The upstream primer contains four tandemcopies of the NF-κB binding site (GGGGACTTTCCC) (SEQ ID NO:8), 18 bp ofsequence complementary to the 5′ end of the SV40 early promotersequence, and is flanked with an XhoI site:5′:GCGGCCTCGAGGGGACTTTCCCGGGGACTTTCCG (SEQ ID NO:9)GGGACTTTCCGGGACTTTCCATCCTGCCATCTCAATT AG:3′

[0559] The downstream primer is complementary to the 3′ end of the SV40promoter and is flanked with a Hind III site:

[0560] 5′:GCGGCAAGCTTTTTGCAAAGCCTAGGC:3′ (SEQ ID NO:4)

[0561] PCR amplification is performed using the SV40 promoter templatepresent in the pB-gal:promoter plasmid obtained from Clontech. Theresulting PCR fragment is digested with XhoI and Hind III and subclonedinto BLSK2-. (Stratagene) Sequencing with the T7 and T3 primers confirmsthe insert contains the following sequence:5′:CTCGAGGGGACTTTCCCGGGGACTTTCCGGGGA (SEQ ID NO:10)CTTTCCGGGACTTTCCATCTGCCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAA AAAGCTT:3′

[0562] Next, replace the SV40 minimal promoter element present in thepSEAP2-promoter plasmid (Clontech) with this NF-κB/SV40 fragment usingXhoI and HindIII. However, this vector does not contain a neomycinresistance gene, and therefore, is not preferred for mammalianexpression systems.

[0563] In order to generate stable mammalian cell lines, theNF-κB/SV40/SEAP cassette is removed from the above NF-κB/SEAP vectorusing restriction enzymes SalI and NotI, and inserted into a vectorcontaining neomycin resistance. Particularly, the NF-κB/SV40/SEAPcassette was inserted into pGFP-1 (Clontech), replacing the GFP gene,after restricting pGFP-1 with SalI and NotI.

[0564] Once NF-κB/SV40/SEAP/Neo vector is created, stable Jurkat T-cellsare created and maintained according to the protocol described inExample 13. Similarly, the method for assaying supernatants with thesestable Jurkat T-cells is also described in example 13. As a positivecontrol, exogenous TNF alpha (0.1,1, 10 ng) is added to wells H9, H10,and H11, with a 5-10 fold activation typically observed.

Example 17 Assay for SEAP Activity

[0565] As a reporter molecule for the assays described in Examples13-16, SEAP activity is assayed using the Tropix Phospho-light Kit (Cat.BP-400) according to the following general procedure. The TropixPhospho-light Kit supplies the Dilution, Assay, and Reaction Buffersused below.

[0566] Prime a dispenser with the 2.5×Dilution Buffer and dispense 15 μlof 2.5×dilution buffer into Optiplates containing 35 μl of asupernatant. Seal the plates with a plastic sealer and incubate at 65°C. for 30 min. Separate the Optiplates to avoid uneven heating.

[0567] Cool the samples to room temperature for 15 minutes. Empty thedispenser and prime with the Assay Buffer. Add 50 μl Assay Buffer andincubate at room temperature 5 min. Empty the dispenser and prime withthe Reaction Buffer (see the table below). Add 50 μl Reaction Buffer andincubate at room temperature for 20 minutes. Since the intensity of thechemiluminescent signal is time dependent, and it takes about 10 minutesto read 5 plates on luminometer, one should treat 5 plates at each timeand start the second set 10 minutes later.

[0568] Read the relative light unit in the luminometer. Set H12 asblank, and print the results. An increase in chemiluminescence indicatesreporter activity. Reaction Buffer Formulation: # of plates Rxn bufferdiluent (ml) CSPD (ml) 10 60 3 11 65 3.25 12 70 3.5 13 75 3.75 14 80 415 85 4.25 16 90 4.5 17 95 4.75 18 100 5 19 105 5.25 20 110 5.5 21 1155.75 22 120 6 23 125 6.25 24 130 6.5 25 135 6.75 26 140 7 27 145 7.25 28150 7.5 29 155 7.75 30 160 8 31 165 8.25 32 170 8.5 33 175 8.75 34 180 935 185 9.25 36 190 9.5 37 195 9.75 38 200 10 39 205 10.25 40 210 10.5 41215 10.75 42 220 11 43 225 11.25 44 230 11.5 45 235 11.75 46 240 12 47245 12.25 48 250 12.5 49 255 12.75 50 260 13

Example 18 High-Throughput Screening Assay Identifying Changes in SmallMolecule Concentration and Membrane Permeability

[0569] Binding of a ligand to a receptor is known to alter intracellularlevels of small molecules, such as calcium, potassium, sodium, and pH,as well as alter membrane potential. These alterations can be measuredin an assay to identify supernatants which bind to receptors of aparticular cell. Although the following protocol describes an assay forcalcium, this protocol can easily be modified to detect changes inpotassium, sodium, pH, membrane potential, or any other small moleculewhich is detectable by a florescent probe.

[0570] The following assay uses Fluorometric Imaging Plate Reader(“FLIPR”) to measure changes in fluorescent molecules (Molecular Probes)that bind small molecules. Clearly, any fluorescent molecule detecting asmall molecule can be used instead of the calcium fluorescent molecule,fluo-3, used here.

[0571] For adherent cells, seed the cells at 10,000-20,000 cells/well ina Co-star black 96-well plate with clear bottom. The plate is incubatedin a CO₂ incubator for 20 hours. The adherent cells are washed two timesin Biotek washer with 200 ul of HBSS (Hank's Balanced Salt Solution)leaving 100 ul of buffer after the final wash.

[0572] A stock solution of 1 mg/ml fluo-3 is made in 10% pluronic acidDMSO. To load the cells with fluo-3, 50 ul of 12 ug/ml fluo-3 is addedto each well. The plate is incubated at 37° C. in a CO₂ incubator for 60min. The plate is washed four times in the Biotek washer with HBSSleaving 100 ul of buffer.

[0573] For non-adherent cells, the cells are spun down from culturemedia. Cells are re-suspended to 2-5×10⁶ cells/ml with l HBSS in a 50-mlconical tube. 4 ul of 1 mg/ml fluo-3 solution in 10% pluronic acid DMSOis added to each ml of cell suspension. The tube is then placed in a 37°C. water bath for 30-60 min. The cells are washed twice with HBSS,resuspended to 1×10⁶ cells/ml, and dispensed into a microplate, 100ul/well. The plate is centrifuged at 1000 rpm for 5 min. The plate isthen washed once in Denley CellWash with 200 ul, followed by anaspiration step to 100 ul final volume.

[0574] For a non-cell based assay, each well contains a fluorescentmolecule, such as fluo-3. The supernatant is added to the well, and achange in fluorescence is detected.

[0575] To measure the fluorescence of intracellular calcium, the FLIPRis set for the following parameters: (1) System gain is 300-800 mW; (2)Exposure time is 0.4 second; (3) Camera F/stop is F/2; (4) Excitation is488 nm; (5) Emission is 530 nm; and (6) Sample addition is 50 ul.Increased emission at 530 nm indicates an extracellular signaling eventwhich has resulted in an increase in the intracellularCa⁺⁺concentration.

Example 19 High-Throughput Screening Assay Identifying Tyrosine KinaseActivity

[0576] The Protein Tyrosine Kinases (PTK) represent a diverse group oftransmembrane and cytoplasmic kinases. Within the Receptor ProteinTyrosine Kinase RPTK) group are receptors for a range of mitogenic andmetabolic growth factors including the PDGF, FGF, EGF, NGF, HGF andInsulin receptor subfamilies. In addition there are a large family ofRPTKs for which the corresponding ligand is unknown. Ligands for RPTKsinclude mainly secreted small proteins, but also membrane-bound andextracellular matrix proteins.

[0577] Activation of RPTK by ligands involves ligand-mediated receptordimerization, resulting in transphosphorylation of the receptor subunitsand activation of the cytoplasmic tyrosine kinases. The cytoplasmictyrosine kinases include receptor associated tyrosine kinases of thesrc-family (e.g., src, yes, lck, lyn, fyn) and non-receptor linked andcytosolic protein tyrosine kinases, such as the Jak family, members ofwhich mediate signal transduction triggered by the cytokine superfamilyof receptors (e.g., the Interleukins, Interferons, GM-CSF, and Leptin).

[0578] Because of the wide range of known factors capable of stimulatingtyrosine kinase activity, the identification of novel human secretedproteins capable of activating tyrosine kinase signal transductionpathways are of interest. Therefore, the following protocol is designedto identify those novel human secreted proteins capable of activatingthe tyrosine kinase signal transduction pathways.

[0579] Seed target cells (e.g., primary keratinocytes) at a density ofapproximately 25,000 cells per well in a 96 well Loprodyne Silent ScreenPlates purchased from Nalge Nunc (Naperville, Ill.). The plates aresterilized with two 30 minute rinses with 100% ethanol, rinsed withwater and dried overnight. Some plates are coated for 2 hr with 100 mlof cell culture grade type I collagen (50 mg/ml), gelatin (2%) orpolylysine (50 mg/ml), all of which can be purchased from SigmaChemicals (St. Louis, Mo.) or 10% Matrigel purchased from BectonDickinson (Bedford,Mass.), or calf serum, rinsed with PBS and stored at4° C. Cell growth on these plates is assayed by seeding 5,000 cells/wellin growth medium and indirect quantitation of cell number through use ofalamarBlue as described by the manufacturer Alamar Biosciences, Inc.(Sacramento, Calif.) after 48 hr. Falcon plate covers #3071 from BectonDickinson (Bedford,Mass.) are used to cover the Loprodyne Silent ScreenPlates. Falcon Microtest III cell culture plates can also be used insome proliferation experiments.

[0580] To prepare extracts, A431 cells are seeded onto the nylonmembranes of Loprodyne plates (20,000/200 ml/well) and culturedovernight in complete medium. Cells are quiesced by incubation inserum-free basal medium for 24 hr. After 5-20 minutes treatment with EGF(60 ng/ml) or 50 ul of the supernatant produced in Example 11, themedium was removed and 100 ml of extraction buffer ((20 mM HEPES pH 7.5,0.15 M NaCl, 1% Triton X-100, 0.1% To SDS, 2 mM Na3VO4, 2 mM Na4P2O7 anda cocktail of protease inhibitors (#1836170) obtained from BoeheringerMannheim (Indianapolis, Ind.) is added to each well and the plate isshaken on a rotating shaker for 5 minutes at 4° C. The plate is thenplaced in a vacuum transfer manifold and the extract filtered throughthe 0.45 mm membrane bottoms of each well using house vacuum. Extractsare collected in a 96-well catch/assay plate in the bottom of the vacuummanifold and immediately placed on ice. To obtain extracts clarified bycentrifugation, the content of each well, after detergent solubilizationfor 5 minutes, is removed and centrifuged for 15 minutes at 4° C. at16,000×g.

[0581] Test the filtered extracts for levels of tyrosine kinaseactivity. Although many methods of detecting tyrosine kinase activityare known, one method is described here.

[0582] Generally, the tyrosine kinase activity of a supernatant isevaluated by determining its ability to phosphorylate a tyrosine residueon a specific substrate (a biotinylated peptide). Biotinylated peptidesthat can be used for this purpose include PSK1 (corresponding to aminoacids 6-20 of the cell division kinase cdc2-p34) and PSK2 (correspondingto amino acids 1-17 of gastrin). Both peptides are substrates for arange of tyrosine kinases and are available from Boehrin ger Mannheim.

[0583] The tyrosine kinase reaction is set up by adding the followingcomponents in order. First, add 10 ul of 5 uM Biotinylated Peptide, then10 ul ATP/Mg₂₊ (5 mM ATP/50 mM MgCl₂), then 10 ul of 5×Assay Buffer (40mM imidazole hydrochloride, pH7.3, 40 nM beta-glycerophosphate, 1 mMEGTA, 100 mM MgCl₂, 5 mM MnCl₂, 0.5 mg/ml BSA), then 5 ul of SodiumVanadate(1 mM), and then 5 ul of water. Mix the components gently andpreincubate the reaction mix at 30° C. for 2 min. Initial the reactionby adding 10 ul of the control enzyme or the filtered supernatant.

[0584] The tyrosine kinase assay reaction is then terminated by adding10 ul of 120 mm EDTA and place the reactions on ice.

[0585] Tyrosine kinase activity is determined by transferring 50 ulaliquot of reaction mixture to a microtiter plate (MTP) module andincubating at 37° C. for 20 min. This allows the streptavadin coated 96well plate to associate with the biotinylated peptide. Wash the MTPmodule with 300 ul/well of PBS four times. Next add 75 ul ofanti-phospotyrosine antibody conjugated to horse radishperoxidase(anti-P-Tyr-POD(0.5 u/ml)) to each well and incubate at 37° C.for one hour. Wash the well as above.

[0586] Next add 100 ul of peroxidase substrate solution (BoehringerMannheim) and incubate at room temperature for at least 5 mins (up to 30min). Measure the absorbance of the sample at 405 nm by using ELISAreader. The level of bound peroxidase activity is quantitated using anELISA reader and reflects the level of tyrosine kinase activity.

Example 20 High-Throughput Screening Assay Identifying PhosphorylationActivity

[0587] As a potential alternative and/or compliment to the assay ofprotein tyrosine kinase activity described in Example 19, an assay whichdetects activation (phosphorylation) of major intracellular signaltransduction intermediates can also be used. For example, as describedbelow one particular assay can detect tyrosine phosphorylation of theErk-1 and Erk-2 kinases. However, phosphorylation of other molecules,such as Raf, JNK, p38 MAP, Map kinase kinase (MEK), MEK kinase, Src,Muscle specific kinase (MuSK), IRAK, Tec, and Janus, as well as anyother phosphoserine, phosphotyrosine, or phosphothreonine molecule, canbe detected by substituting these molecules for Erk-1 or Erk-2 in thefollowing-assay.

[0588] Specifically, assay plates are made by coating the wells of a96-well ELISA plate with 0.1 ml of protein G (1 ug/ml) for 2 hr at roomtemp, (RT). The plates are then rinsed with PBS and blocked with 3%BSA/PBS for 1 hr at RT. The protein G plates are then treated with 2commercial monoclonal antibodies (100 ng/well) against Erk-1 and Erk-2(1 hr at RT) (Santa Cruz Biotechnology). (To detect other molecules,this step can easily be modified by substituting a monoclonal antibodydetecting any of the above described molecules.) After 3-5 rinses withPBS, the plates are stored at 4° C. until use.

[0589] A431 cells are seeded at 20,000/well in a 96-well Loprodynefilterplate and cultured overnight in growth medium. The cells are thenstarved for 48 hr in basal medium (DNEM) and then treated with EGF (6ng/well) or 50 ul of the supernatants obtained in Example 11 for 5-20minutes. The cells are then solubilized and extracts filtered directlyinto the assay plate.

[0590] After incubation with the extract for 1 hr at RT, the wells areagain rinsed. As a positive control, a commercial preparation of MAPkinase (10 ng/well) is used in place of A431 extract. Plates are thentreated with a commercial polyclonal (rabbit) antibody (1 ug/ml) whichspecifically recognizes the phosphorylated epitope of the Erk-1 andErk-2 kinases (1 hr at RT). This antibody is biotinylated by standardprocedures. The bound polyclonal antibody is then quantitated bysuccessive incubations with Europium-streptavidin and Europiumfluorescence enhancing reagent in the Wallac DELFIA instrument(time-resolved fluorescence). An increased fluorescent signal overbackground indicates a phosphorylation.

Example 21 Method of Determining Alterations in a Gene Corresponding toa Polynucleotide

[0591] RNA isolated from entire families or individual patientspresenting with a phenotype of interest (such as a disease) is beisolated. cDNA is then generated from these RNA samples using protocolsknown in the art. (See, Sambrook.) The cDNA is then used as a templatefor PCR, employing primers surrounding regions of interest in SEQ IDNO:X. Suggested PCR conditions consist of 35 cycles at 95° C. for 30seconds; 60-120 seconds at 52-58° C.; and 60-120 seconds at 70° C.,using buffer solutions described in Sidransky, D., et al., Science252:706 (1991).

[0592] PCR products are then sequenced using primers labeled at their 5′end with T4 polynucleotide kinnase, employing SequiTherm Polymerase.(Epicentre Technologies). The intron-exon borders of selected exons isalso determined and genomic PCR products analyzed to confirm theresults. PCR products harboring suspected mutations is then cloned andsequenced to validate the results of the direct sequencing .

[0593] PCR products is cloned into T-tailed vectors as described inHolton, T. A. and Graham, M. W., Nucleic Acids Research, 19:1156 (1991)and sequenced with T7 polymerase (United States Biochemical). Affectedindividuals are identified by mutations not present in unaffectedindividuals.

[0594] Genomic rearrangements are also observed as a method ofdetermining alterations in a gene corresponding to a polynucleotide.Genomic clones isolated according to Example 2 are nick-translated withdigoxigenindeoxy-uridine 5′-triphosphate (Boehringer Manheim), and FISHperformed as described in Johnson, Cg. et al., Methods Cell Biol.35:73-99 (1991). Hybridization with the labeled probe is carried outusing a vast excess of human cot-1 DNA for specific hybridization to thecorresponding genomic locus.

[0595] Chromosomes are counterstained with 4,6-diamino-2-phenylidole andpropidium iodide, producing a combination of C- and R-bands. Alignedimages for precise mapping are obtained using a triple-band filter set(Chroma Technology, Brattleboro, Vt.) in combination with a cooledcharge-coupled device camera (Photometrics, Tucson, Ariz.) and variableexcitation wavelength filters. (Johnson, Cv. et al., Genet. Anal. Tech.Appl., 8:75 (1991).) Image collection, analysis and chromosomalfractional length measurements are performed using the ISee GraphicalProgram System. (Inovision Corporation, Durham, N.C.) Chromosomealterations of the genomic region hybridized by the probe are identifiedas insertions, deletions, and translocations. These alterations are usedas a diagnostic marker for an associated disease.

Example 22 Method of Detecting, Abnormal Levels of a Polypeptide in aBiological Sample

[0596] A polypeptide of the present invention can be detected in abiological sample, and if an increased or decreased level of thepolypeptide is detected, this polypeptide is a marker for a particularphenotype. Methods of detection are numerous, and thus, it is understoodthat one skilled in the art can modify the following assay to fit theirparticular needs.

[0597] For example, antibody-sandwich ELISAs are used to detectpolypeptides in a sample, preferably a biological sample. Wells of amicrotiter plate are coated with specific antibodies, at a finalconcentration of 0.2 to 10 ug/ml. The antibodies are either monoclonalor polyclonal and are produced by the method described in Example 10.The wells are blocked so that non-specific binding of the polypeptide tothe well is reduced.

[0598] The coated wells are then incubated for >2 hours at RT with asample containing the polypeptide. Preferably, serial dilutions of thesample should be used to validate results. The plates are then washedthree times with deionized or distilled water to remove unboundedpolypeptide.

[0599] Next, 50 ul of specific antibody-alkaline phosphatase conjugate,at a concentration of 25-400 ng, is added and incubated for 2 hours atroom temperature. The plates are again washed three times with deionizedor distilled water to remove unbounded conjugate.

[0600] Add 75 ul of 4-methylumbelliferyl phosphate (MUP) orp-nitrophenyl phosphate (NPP) substrate solution to each well andincubate 1 hour at room temperature. Measure the reaction by amicrotiter plate reader. Prepare a standard curve, using serialdilutions of a control sample, and plot polypeptide concentration on theX-axis (log scale) and fluorescence or absorbance of the Y-axis (linearscale). Interpolate the concentration of the polypeptide in the sampleusing the standard curve.

Example 23 Formulating a Polypeptide

[0601] The secreted polypeptide composition will be formulated and dosedin a fashion consistent with good medical practice, talking into accountthe clinical condition of the individual patient (especially the sideeffects of treatment with the secreted polypeptide alone), the site ofdelivery, the method of administration, the scheduling ofadministration, and other factors known to practitioners. The “effectiveamount” for purposes herein is thus determined by such considerations.

[0602] As a general proposition, the total pharmaceutically effectiveamount of secreted polypeptide administered parenterally per dose willbe in the range of about 1 μg/kg/day to 10 mg/kg/day of patient bodyweight, although, as noted above, this will be subject to therapeuticdiscretion. More preferably, this dose is at least 0.01 mg/kg/day, andmost preferably for humans between about 0.01 and 1 mg/kg/day for thehormone. If given continuously, the secreted polypeptide is typicallyadministered at a dose rate of about 1 μg/kg/hour to about 50μg/kg/hour, either by 1-4 injections per day or by continuoussubcutaneous infusions, for example, using a mini-pump. An intravenousbag solution may also be employed. The length of treatment needed toobserve changes and the interval following treatment for responses tooccur appears to vary depending on the desired effect.

[0603] Pharmaceutical compositions containing the secreted protein ofthe invention are administered orally, rectally, parenterally,intracistemally, intravaginally, intraperitoneally, topically (as bypowders, ointments, gels, drops or transdermal patch), bucally, or as anoral or nasal spray. “Pharmaceutically acceptable carrier” refers to anon-toxic solid, semisolid or liquid filler, diluent, encapsulatingmaterial or formulation auxiliary of any type. The term “parenteral” asused herein refers to modes of administration which include intravenous,intramuscular, intraperitoneal, intrastemal, subcutaneous andintraarticular injection and infusion.

[0604] The secreted polypeptide is also suitably administered bysustained-release systems. Suitable examples of sustained-releasecompositions include semi-permeable polymer matrices in the form ofshaped articles, e.g., films, or mirocapsules. Sustained-releasematrices include polylactides (U.S. Pat. No. 3,773,919, EP 58,481),copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman, U. etal., Biopolymers 22:547-556 (1983)), poly (2- hydroxyethyl methacrylate)(R. Langer et al., J. Biomed. Mater. Res. 15:167-277 (1981), and R.Langer, Chem. Tech. 12:98-105 (1982)), ethylene vinyl acetate (R. Langeret al.) or poly-D- (−)-3-hydroxybutyric acid (EP 133,988).Sustained-release compositions also include liposomally entrappedpolypeptides. Liposomes containing the secreted polypeptide are preparedby methods known per se: DE 3,218,121; Epstein et al., Proc. Natl. Acad.Sci. USA 82:3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA77:4030-4034 (1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP142,641; Japanese Pat. Appl. 83-118008; U.S. Pat. Nos. 4,485,045 and4,544,545; and EP 102,324. Ordinarily, the liposomes are of the small(about 200-800 Angstroms) unilamellar type in which the lipid content isgreater than about 30 mol. percent cholesterol, the selected proportionbeing adjusted for the optimal secreted polypeptide therapy.

[0605] For parenteral administration, in one embodiment, the secretedpolypeptide is formulated generally by mixing it at the desired degreeof purity, in a unit dosage injectable form (solution, suspension, oremulsion), with a pharmaceutically acceptable carrier, i.e., one that isnon-toxic to recipients at the dosages and concentrations employed andis compatible with other ingredients of the formulation. For example,the formulation preferably does not include oxidizing agents and othercompounds that are known to be deleterious to polypeptides.

[0606] Generally, the formulations are prepared by contacting thepolypeptide uniformily and intimately with liquid carriers or finelydivided solid carriers or both. Then, if necessary, the product isshaped into the desired formulation. Preferably the carrier is aparenteral carrier, more preferably a solution that is isotonic with theblood of the recipient. Examples of such carrier vehicles include water,saline, Ringer's solution, and dextrose solution. Non-aqueous vehiclessuch as fixed oils and ethyl oleate are also useful herein, as well asliposomes.

[0607] The carrier suitably contains minor amounts of additives such assubstances that enhance isotonicity and chemical stability. Suchmaterials are non-toxic to recipients at the dosages and concentrationsemployed, and include buffers such as phosphate, citrate, succinate,acetic acid, and other organic acids or their salts; antioxidants suchas ascorbic acid; low molecular weight (less than about ten residues)polypeptides, e.g., potyarginine or tripeptides; proteins, such as serumalbumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; a imino acids, such as glycine, glutamic acid,aspartic acid, or arginine; monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, manose,or dextrins; chelating agents such as EDTA; sugar alcohols such asmannitol or sorbitol; counterions such as sodium; and/or nonionicsurfactants such as polysorbates, poloxamers, or PEG.

[0608] The secreted polypeptide is typically formulated in such vehiclesat a concentration of about 0.1 mg/ml to 100 mg/ml, preferably 1-10mg/ml, at a pH of about 3 to 8. It will be understood that the use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of polypeptide salts.

[0609] Any polypeptide to be used for therapeutic administration can besterile. Sterility is readily accomplished by filtration through sterilefiltration membranes (e.g., 0.2 micron membranes). Therapeuticpolypeptide compositions generally are placed into a container having asterile access port, for example, an intravenous solution bag or vialhaving a stopper pierceable by a hypodermic injection needle.

[0610] Polypeptides ordinarily will be stored in unit or multi-dosecontainers, for example, sealed ampoules or vials, as an aqueoussolution or as a lyophilized formulation for reconstitution. As anexample of a lyophilized formulation, 10-ml vials are filled with 5 mlof sterile-filtered 1% (w/v) aqueous polypeptide solution, and theresulting mixture is lyophilized. The infusion solution is prepared byreconstituting the lyophilized polypeptide using bacteriostaticWater-for-Injection.

[0611] The invention also provides a pharmaceutical pack or kitcomprising one or more containers filled with one or more of theingredients of the pharmaceutical compositions of the invention.Associated with such container(s) can be a notice in the form prescribedby a governmental agency regulating the manufacture, use or sale ofpharmaceuticals or biological products, which notice reflects approvalby the agency of manufacture, use or sale for human administration. Inaddition, the polypeptides of the present invention may be employed inconjunction with other therapeutic compounds.

Example 24 Method of Treating Decreased Levels of the Polypeptide

[0612] It will be appreciated that conditions caused by a decrease inthe standard or normal expression level of a secreted protein in anindividual can be treated by administering the polypeptide of thepresent invention, preferably in the secreted form. Thus, the inventionalso provides a method of treatment of an individual in need of anincreased level of the polypeptide comprising administering to such anindividual a pharmaceutical composition comprising an amount of thepolypeptide to increase the activity level of the polypeptide in such anindividual.

[0613] For example, a patient with decreased levels of a polypeptidereceives a daily dose 0.1-100 ug-kg of the polypeptide for sixconsecutive days. Preferably, the polypeptide is in the secreted form.The exact details of the dosing scheme, based on administration andformulation, are provided in Example 23.

Example 25 Method of Treating Increased Levels of the Polypeptide

[0614] Antisense technology is used to inhibit production of apolypeptide of the present invention. This technology is one example ofa method of decreasing levels of a polypeptide, preferably a secretedform, due to a variety of etiologies, such as cancer.

[0615] For example, a patient diagnosed with abnormally increased levelsof a polypeptide is administered intravenously antisense polynucleotidesat 0.5, 1.0, 1.5, 2.0 and 3.0 mg/kg day for 21 days. This treatment isrepeated after a 7-day rest period if the treatment was well tolerated.The formulation of the antisense polynucleotide is provided in Example23.

Example 26 Method of Treatment Using Gene Therapy

[0616] One method of gene therapy transplants fibroblasts, which arecapable of expressing a polypeptide, onto a patient. Generally,fibroblasts are obtained from a subject by skin biopsy. The resultingtissue is placed in tissue-culture medium and separated into smallpieces. Small chunks of the tissue are placed on a wet surface of atissue culture flask, approximately ten pieces are placed in each flask.The flask is turned upside down, closed tight and left at roomtemperature over night. After 24 hours at room temperature, the flask isinverted and the chunks of tissue remain fixed to the bottom of theflask and fresh media (e.g., Ham's F12 media, with 10% FBS, penicillinand streptomycin) is added. The flasks are then incubated at 37° C. forapproximately one week.

[0617] At this time, fresh media is added and subsequently changed everyseveral days. After an additional two weeks in culture, a monolayer offibroblasts emerge. The monolayer is trypsinized and scaled into largerflasks.

[0618] pMV-7 (Kirschmeier, P. T. et al., DNA, 7:219-25 (1988)), flankedby the long terminal repeats of the Moloney murine sarcoma virus, isdigested with EcoRI and HindIII and subsequently treated with calfintestinal phosphatase. The linear vector is fractionated on agarose geland purified, using glass beads.

[0619] The cDNA encoding a polypeptide of the present invention can beamplified using PCR primers which correspond to the 5′ and 3′ endsequences respectively as set forth in Example 1. Preferably, the 5′primer contains an EcoRI site and the 3′ primer includes a HindIII site.Equal quantities of the Moloney murine sarcoma virus linear backbone andthe amplified EcoRI and HindIII fragment are added together, in thepresence of T4 DNA ligase. The resulting mixture is maintained underconditions appropriate for ligation of the two fragments. The ligationmixture is then used to transform bacteria HB101, which are then platedonto agar containing kanamycin for the purpose of confirming that thevector has the gene of interest properly inserted.

[0620] The amphotropic pA317 or GP+am12 packaging cells are grown intissue culture to confluent density in Dulbecco's Modified Eagles Medium(DMEM) with 10% calf serum (CS), penicillin and streptomycin. The MSVvector containing the gene is then added to the media and the packagingcells transduced with the vector. The packaging cells now produceinfectious viral particles containing the gene (the packaging cells arenow referred to as producer cells).

[0621] Fresh media is added to the transduced producer cells, andsubsequently, the media is harvested from a 10 cm plate of confluentproducer cells. The spent media, containing the infectious viralparticles, is filtered through a millipore filter to remove detachedproducer cells and this media is then used to infect fibroblast cells.Media is removed from a sub-confluent plate of fibroblasts and quicklyreplaced with the media from the producer cells. This media is removedand replaced with fresh media. If the titer of virus is high, thenvirtually all fibroblasts will be infected and no selection is required.If the titer is very low, then it is necessary to use a retroviralvector that has a selectable marker, such as neo or his. Once thefibroblasts have been efficiently infected, the fibroblasts are analyzedto determine whether protein is produced.

[0622] The engineered fibroblasts are then transplanted onto the host,either alone or after having been grown to confluence on cytodex 3microcarrier beads.

[0623] It will be clear that the invention may be practiced otherwisethan as particularly described in the foregoing description andexamples. Numerous modifications and variations of the present inventionare possible in light of the above teachings and, therefore, are withinthe scope of the appended claims.

[0624] The entire disclosure of each document cited (including patents,patent applications, journal articles, abstracts, laboratory manuals,books, or other disclosures) in the Background of the Invention,Detailed Description, and Examples is hereby incorporated herein byreference. Further, the hard copy of the sequence listing submittedherewith and the corresponding computer readable form are bothincorporated herein by reference in their entireties.

1 229 1 733 DNA Homo sapiens 1 gggatccgga gcccaaatct tctgacaaaactcacacatg cccaccgtgc ccagcacctg 60 aattcgaggg tgcaccgtca gtcttcctcttccccccaaa acccaaggac accctcatga 120 tctcccggac tcctgaggtc acatgcgtggtggtggacgt aagccacgaa gaccctgagg 180 tcaagttcaa ctggtacgtg gacggcgtggaggtgcataa tgccaagaca aagccgcggg 240 aggagcagta caacagcacg taccgtgtggtcagcgtcct caccgtcctg caccaggact 300 ggctgaatgg caaggagtac aagtgcaaggtctccaacaa agccctccca acccccatcg 360 agaaaaccat ctccaaagcc aaagggcagccccgagaacc acaggtgtac accctgcccc 420 catcccggga tgagctgacc aagaaccaggtcagcctgac ctgcctggtc aaaggcttct 480 atccaagcga catcgccgtg gagtgggagagcaatgggca gccggagaac aactacaaga 540 ccacgcctcc cgtgctggac tccgacggctccttcttcct ctacagcaag ctcaccgtgg 600 acaagagcag gtggcagcag gggaacgtcttctcatgctc cgtgatgcat gaggctctgc 660 acaaccacta cacgcagaag agcctctccctgtctccggg taaatgagtg cgacggccgc 720 gactctagag gat 733 2 5 PRT Homosapiens Site (3) Xaa equals any of the twenty naturally ocurring L-aminoacids 2 Trp Ser Xaa Trp Ser 1 5 3 86 DNA Homo sapiens 3 gcgcctcgagatttccccga aatctagatt tccccgaaat gatttccccg aaatgatttc 60 cccgaaatatctgccatctc aattag 86 4 27 DNA Homo sapiens 4 gcggcaagct ttttgcaaagcctaggc 27 5 271 DNA Homo sapiens 5 ctcgagattt ccccgaaatc tagatttccccgaaatgatt tccccgaaat gatttccccg 60 aaatatctgc catctcaatt agtcagcaaccatagtcccg cccctaactc cgcccatccc 120 gcccctaact ccgcccagtt ccgcccattctccgccccat ggctgactaa ttttttttat 180 ttatgcagag gccgaggccg cctcggcctctgagctattc cagaagtagt gaggaggctt 240 ttttggaggc ctaggctttt gcaaaaagct t271 6 32 DNA Homo sapiens 6 gcgctcgagg gatgacagcg atagaacccc gg 32 7 31DNA Homo sapiens 7 gcgaagcttc gcgactcccc ggatccgcct c 31 8 12 DNA Homosapiens 8 ggggactttc cc 12 9 73 DNA Homo sapiens 9 gcggcctcga ggggactttcccggggactt tccggggact ttccgggact ttccatcctg 60 ccatctcaat tag 73 10 256DNA Homo sapiens 10 ctcgagggga ctttcccggg gactttccgg ggactttccgggactttcca tctgccatct 60 caattagtca gcaaccatag tcccgcccct aactccgcccatcccgcccc taactccgcc 120 cagttccgcc cattctccgc cccatggctg actaattttttttatttatg cagaggccga 180 ggccgcctcg gcctctgagc tattccagaa gtagtgaggaggcttttttg gaggcctagg 240 cttttgcaaa aagctt 256 11 1169 DNA Homo sapiensSITE (1151) n equals a,t,g, or c 11 ggggcgcaaa tagggtcagt gggccgcttggcgktgttcg ttgcggtacc aggtccgcgt 60 gaggggttcg ggggttctgg gcaggcacaatggcgtctcg agcaggcccg cgagcggccg 120 rcaccgacgc agcgagcttt cagcaccgggagcgcgtcgc catgcactac cagatgagtg 180 tgaccctcaa gtatgaaatc aagaagctgatctacgtaca tctggtcata tggctgctgc 240 tggttgctaa gatgagcgtg ggacacctgaggctcttgtc acatgatcag gtggccatgc 300 cctatcagtg ggaatacccg tatttgctgagcattttgcc ctctctcttg ggccttctct 360 cctttccccg caacaacatt agctacctggtgctctccat gatcagcatg ggactctttt 420 ccatcgctcc actcatttat ggcagcatggagatgttccc tgctgcacag ccttctaccg 480 ccatggcaag gcctaccgtt tcctctttggtttttctgcc gtttccatca tgtacctggt 540 gttggtgttg gcagtgcaag tgcatgcctggcagttgtac tacagcaaga agctcctaga 600 ctcttggttc accagcacac aggagaagaagcataaatga agcctctttg gggtgaagcc 660 tggacatccc atcgaatgaa aggacactagtacagcggtt ccaaaatccc ttctggtgat 720 tttagcagct gtgatgttgg tacctggtgcagacccaggc caaagttctg gaaagctcct 780 tttgccatct gctgaggtgg caaaactataatttattcct ggttggctag aactgggtga 840 ccaacagcta tgaaacaaat ttcagctgtttgaagttgaa ctttgaggtt tttctttaag 900 aatgagcttc gtccttgcct ctactcggtcattctcccca tttccatcca ttacccctta 960 gccattgaga ctaaaggaaa tagggaataaatcaaattac ttcatctcta ggtcacgggt 1020 caggaaacat ttgggcagct gctcccttggcagctgtggt ctcctctgca aagcatttta 1080 attaaaaacc tcaataaaga tgccctgcccacaaaaaaaa aaaaaaaaaa aattcggggg 1140 ggggcccggg naaccaattn gcccctana1169 12 1310 DNA Homo sapiens 12 aattcggcac gaggcagcgt cgcgcggcccagttcccttt tccggtcggc gtggtcttgc 60 gagtggagtg tccgctgtgc ccgggcctgcaccatgagcg tcccggcctt catcgacatc 120 agtgaagaag atcaggctgc tgagcttcgtgcttatctga aatctaaagg agctgagatt 180 tcagaagaga actcggaagg tggacttcatgttgatttag ctcaaattat tgaagcctgt 240 gatgtgtgtc tgaaggagga tgataaagatgttgaaagtg tgatgaacag tgtggtatcc 300 ctactcttga tcctggaacc agacaagcaagaagctttga ttgaaagcct atgtgaaaag 360 ctggtcaaat ttcgcgaagg tgaacgcccgtctctgagac tgcagttgtt aagcaacctt 420 ttccacggga tggataagaa tactcctgtaagatacacag tgtattgcag ccttattaaa 480 gtggcagcat cttgtggggc catccagtacatcccaactg agctggatca agttagaaaa 540 tggatttctg actggaatct caccactgaaaaaaagcaca cccttttaag actactttat 600 gaggcacttg tggattgtaa gaagagtgatgctgcttcaa aagtcatggt ggaattgctc 660 ggaagttaca cagaggacaa tgcttcccaggctcgagttg atgcccacag gtgtattgta 720 cgagcattga aagatccaaa tgcatttctttttgaccacc ttcttacttt aaaaccagtc 780 aagtttttgg aaggcgagct tattcatgatcttttaacca tttttgtgag tgctaaattg 840 gcatcatatg tcaagtttta tcagaataataaagacttca ttgattcact tggcctgtta 900 catgaacaga atatggcaaa aatgagactacttactttta tgggaatggc agtagaaaat 960 aaggaaattt cttttgacac aatgcagcaagaacttcaga ttggagctga tgatgttgaa 1020 gcatttgtta ttgacgccgt aagaactaaaatggtctact gcaaaattga tcagacccag 1080 agaaaagtag ttgtcagtca tagcacacatcggacatttg gaaaacagca gtggcaacaa 1140 ctgtatgaca cacttaatgc ctggaaacaaaatctgaaca aagtgaaaaa cagccttttg 1200 agtctttctg atacctgagt ttttatgcttataatttttg ttctttgaaa aaaaagccct 1260 aaatcatagt aaaacattat aaactaaaaaaaaaaaaaaa aaaaaaaaaa 1310 13 1139 DNA Homo sapiens SITE (7) n equalsa,t,g, or c 13 agggcanact tacagagata tcatatgaga tcacccctcg cattcgtgtctggcgccaga 60 ccctcgagcg gtgccggagc gcasccaggt gtgcttgtgc ctgggccagctggagaggtc 120 cattgcctgg gangaagtct gtcaacaaag tgacatgtct agtctgccggaagggtgaca 180 atgatgagtt tcttctgctt tgtgatgggt gtraccgtgg ctgccacatttactgccatc 240 gtcccaagat ggaggctgtc ccagaaggag attggttctg tactgtctgtttggctcagc 300 aggtggaggg agaattcact cagaagcctg gtttcccaaa gcgtggccagaagcggaaaa 360 gtggttattc gctgaacttc tcagagggtg atggccgccg acgccgggtactgttgaggg 420 gccgagaaag cccagcagca gggcctcggt actcggaaga agggctctccccctccaagc 480 ggcggcgact ctctatgcgg aaccaccaca gtgatctcac attttgcgagattatcctga 540 tggagatgga gtcccatgat gcagcctggc ctttcctaga gcctgtgaacccacgtttgg 600 tgagtgggta ccggcgcatc atcaaaaatc ctatggattt ttccaccatgcgggagcggc 660 tgctcagggg agggtacacc agctcagagg agtttgcggc tgatgccctcctggtatttg 720 acaactgcca gactttcaac gaggatgact ctgaagtagg caaggctgggcacatcatgc 780 gccgcttctt cgagagccgc tgggaggagt tttatcaggg aaaacaggccaatctgtgag 840 gcaagggagg tggggagtca ccttgtggca tctcccccca ccttccaaacaaaaacctgc 900 cattttcacc tgctgatgct gccctgggtc cagactcaag tcagatacaaccctgatttt 960 tgaccttncc cttggcagtg ccccacatcc tcttattcct acatccctttctcccttccc 1020 tcctcttgct cctcaagtaa gaggtgcaga gatgaggtcc ttctggactaaaagccaaaa 1080 aaagaaagaa aaaaawaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaaaaaaaan 1139 14 2271 DNA Homo sapiens 14 gttccggggg atgccagctcacttctcgga cagcgcccag actgaggcct gctaccacat 60 gctgagccgg ccccagccgccacccgaccc cctcctgctc cagcgtctgc cacggcccag 120 ctccctgtca gacaagacccagctccacag caggtggctg gactcgtcgc ggtgtctcat 180 gcagcagggc atcaaggccggggacgcact ctggctgcgc ttcaagtact acagcttctt 240 cgatttggat cccaagacagaccccgtgcg gctgacacag ctgtatgagc aggcccggtg 300 ggacctgctg ctggaggagattgactgcac cgaggaggag atgatggtgt ttgccgccct 360 gcagtaccac atcaacaagctgtcccagag cggggaggtg ggggagccgg ctggcacaga 420 cccagggctg gacgacctggatgtggccct gagcaacctg gaggtgaagc tggaggggtc 480 ggcgcccaca gatgtgctggacagcctcac caccatccca gagctcaagg accatctccg 540 aatctttcgg ccccggaagctgaccctgaa gggctaccgc caacactggg tggtgttcaa 600 ggagaccaca ctgtcctactacaagagcca ggacgaggcc cctggggacc ccattcagca 660 gctcaacctc aagggctgtgaggtggttcc cgatgttaac gtctccggcc agaagttctg 720 cattaaactc ctagtgccctcccctgaggg catgagtgag atctacctgc ggtgccagga 780 tgagcagcag tatgcccgctggatggctgg ctgccgcctg gcctccaaag gccgcaccat 840 ggccgacagc agctacaccagcgaggtgca ggccatcctg gccttcctca gcctgcagcg 900 cacgggcagt gggggcccgggcaaccaccc ccacggccct gatgcctctg ccgagggcct 960 caacccctac ggcctcgttgccccccgttt ccagcgaaag ttcaaggcca agcagctcac 1020 cccacggatc ctggaagcccaccagaatgt ggcccagttg tcgctggcag aggcccagct 1080 gcgcttcatc caggcctggcagtccctgcc cgacttcggc atctcctatg tcatggtcag 1140 gttcaagggc agcaggaaagacgagatcct gggcatcgcc aacaaccgac tgatccgcat 1200 cgacttggcc gtgggcgacgtggtcaagac ctggcgtttc agcaacatgc gccagtggaa 1260 tgtcaactgg gacatccggcaggtggccat cgagtttgat gaacacatca atgtggcctt 1320 cagctgcgtg tctgccagctgccgaattgt acacgagtat atcgggggct acattttcct 1380 gtcgacgcgg gagcgggcccgtggggagga gctggatgaa gacctcttcc tgcagctcac 1440 cgggggccat gaggccttctgagggctgtc tgattgcccc tgccctgctc accaccctgt 1500 cacagccact cccaagcccacacccacagg ggctcactgc cccacacccg ctccaggcag 1560 gcacccagct gggcatttcacctgctgtca ctgactttgt gcaggccaag gacctggcag 1620 ggccagacgc tgtaccatcacccaggccag ggatgggggt gggggtccct gagctcatgt 1680 ggtgccccct ttccttgtctgagtggctga ggctgatacc cctgacctat ctgcagtccc 1740 ccagcacaca aggaagaccagatgtagcta caggatgatg aaacatggtt tcaaacgagt 1800 tctttcttgt tactttttaaaatttctttt ttataaatta atattttatt gttggatcct 1860 cctcctttct ctggagctgtgcttggggct actctgacac tctgtctctt catcaccagc 1920 caaggaaagg ggctttcctgataaagacaa gagttggtta gagaaaggga cacctaagtc 1980 agtctagggt tggaagctaggagagaggtg agggcagaag ggcacagctt tcaggaacaa 2040 ggaatagggg ctggggtkgtkgttctcacg ggtaggcgta cctgcagggc ctccttgaag 2100 tacttgggaa ggaggaagccatcagtattc cctggagtca gaatcacccc attggcagag 2160 cggaagaagg gtattccatctgctgacaga gccagagatg tgactcatgc cctccccgaa 2220 ggcaaagtca gctcctgctttgtccagact cacctgccag agccaggggt c 2271 15 626 DNA Homo sapiens SITE(22) n equals a,t,g, or c 15 acaacaaaca tcgaaaatcg antatgtgcc ccgaaaagtcggaacgcagg caatcagtcc 60 gcacgmgcgc aagttcaaca tgaagatgat atgaggccggggcggggggc agggaccccc 120 gggcggccgg gcaggggaag gggcctggcc gccacctgctcactctccag tccttcccac 180 ctcctcccta cccttctaca cacgttctct ttctccctcccgcctccgtc ccctgctgcc 240 ccccgccagc cctcaccacc tgccctcctt ctaccaggacctcagaagcc cagacctggg 300 gaccccacct acacaggggc attgacagac tggagttgaaagccgacgaa ccgacacgcg 360 gcagagtcaa taattcaata aaaaagttac gaactttctctgtaacttgg gtttcaataa 420 ttatggattt ttatgaaaac ttgaaataat aaaaagagaaaaaaactatt tcctatagct 480 agtcggaatg caaacttttg acgtcctgat tgctccagggccctctttcc aactcagttt 540 cttgtttttc ctcttcctcc tcctcctctt cttcctcctttctttctctt nccccatggg 600 ggaggggttc attcagggaa aacagg 626 16 2118 DNAHomo sapiens 16 ttttccagcc atgtcactaa ttgtgaattc ctaccaacta ttgacagaatacagagttga 60 ttttttaata aaaagttata tataattatc cctttaatta aagggagcaaaggggcgttc 120 cacatggaca gaggcttgga ccgaggcctg gtcacagcag cgagcatccagggtttgcag 180 ggacgatgtt acagactctg ttttctgcct ggcgtttcac ttgtgtctgctcctagcctg 240 tgctctgcca gcagcacaga catctgctcc atcagacctc ttccattttgcacagggagt 300 gcaggaggtg aatgttcact ttctgttctc cagtgtcact gttctgtttccacgggatgg 360 aaagcgcatg ggcctgtgtc cattgtagat ttccttctag atttctgtgtacacacactt 420 gattgttctg gatgaatgtc ttttttaata ctccgaaaat ttcatcatctaagaaaatga 480 ttccatacaa ataactcagc acacaagtga cccaggacat atgcctgccaaagggatgtg 540 ttagaaggct gccttctcat gcgcattgtc acttggatct tgtggtgaggacggccccat 600 ctttcttgcc acagattgag gccacttttg agcaagggag atcctggagttaagacaggt 660 gttgggggca gcctgtattt taccctaggg gcaggtctgc atggtgaccccacatygcac 720 tggtaaacca tttgagtccc actcttcatc ctggaagtgg gaactggagtcccacccaca 780 gtgcattcag aaagcatgct gtgtgggggc tgcttctcag gaggccaggcccttctgagc 840 ggaaccgtcc tggagagagc ctgccctcgt ttccaggctg cagccgtaacgcactttctc 900 ccaggctgag ggcgggtgtt ctggggtgtc tgccctctgt cggccctgcttcctgccagg 960 acgtggcctc ttccgatcct tttctctcag acactggagg tctcttctgccattgtgctg 1020 gtcccatccc aagaattgta ggacagagac cacactgggt cggcggacacaaagtccatc 1080 caggacccag gccgcagagg gagcaggaag agatgctgat agtttgatctagaaaccagc 1140 agctactggc tcaaattcag gttctggcgt caaatagcga catttccagtttctcttaaa 1200 aaccgtgttt ggtttcagtt gggataggct tgttttgtct gttgaaaatgtttctagttt 1260 tttttctttc atttttctct cattccattt ctgccttaac tttagtttgttcacagggag 1320 gcaaagctga catgaacctt ttgtcgtggg acttcaggcc acattggcttgaaggcattc 1380 gtttccttct ggggtgggga caggccctca tggcaggctt gttcccgtggctctgagcga 1440 ggcctcttcc tgctgggctc ccagactcct gcatccaggc ccccaccttctcggcttctg 1500 gtttttcttt ctttttggta gaacacaaca tctaccattc agttaaaccttctttatctc 1560 ctcctytggc atccattttt ccaaagaaga gtcgagtcct ctgaggtctgtgcttgaaar 1620 ccgtccgaag gcattcttgt tagctttgct tttctcccca tatcccaaggcgaagcgctg 1680 agattcttcc atctaaaaaa ccctcgaccc gaaaccctca ccagataaactacagtttgt 1740 ttaggaggcc ctgaccttca tggtgtcttt gaagcccaac cactcggtttccttcggatt 1800 ttcctccctt tgttcggggt ttggtttggc tcctctgtgt gtgtccgtatcttgttcggt 1860 gtcctcgagg ttgagcttca ctccactgcg gcagaggcag cgtgcacactcggatttgct 1920 acgtttctat atatcttgaa gctaaatgta tatatgagta gtttgccatgagataacaca 1980 gtgtaaacag tagacaccca gaaatcgtga cttctgtgtt ctctccatttgagtattttg 2040 taattttttt gaaatatttg tggacataaa taaaaccaag ctacactacaaaaaaaaaaa 2100 aaaaaaactg gagactag 2118 17 1076 DNA Homo sapiens SITE(979) n equals a,t,g, or c 17 gcccaaggag ctcagcttcg cccgcatcaaggccgttgag tgcgtggaga gcaccgggcg 60 ccacatctac ttcacgctgg tgaccgaagggwgcggcgag atcgacttcc gctgccccct 120 ggaagatccc ggctggaacg cccagatcaccctaggcctg gtcaagttca agaaccagca 180 ggccatccag acagtgcggg cccggcagagcctcgggacc gggaccctcg tgtcctaaac 240 caccgggcgc accatctttc cttcatgctacccaccacct cagtgctgag gtcaaggcag 300 cttcgttgtt ccctctggct tgtgggggcacggctgtsyt ccatgtggca aggtggaagg 360 catggacgtg tggaggaggc gctggagctgaaggaatgga cgagccctgg gaggagggca 420 gaaggctacg cagggctgag gatgaagatgcagcccctgg atggtcccag actctcagga 480 catgcccagc tcaggggctt cgagccacaggcctggcctc atatggcatg agggggagct 540 ggcataggag ccccctccct gctgtggtcctgccctctgt cctgcagact gctcttagcc 600 ccctggcttt gtgccaggcc tggaggagggcagtccccca tggggtgccg agccaacgcc 660 tcaggaatca ggaggccagc ctggtaccaaaaggagtacc cagggcctgg tacccaggcc 720 cactccagaa tggcctctgg actcaccttgagaaggggga gctgctgggc ctaaagccca 780 ctcctggggg tctcctgctg cttaggtccttttgggaccc ccacccatcc aggccctttc 840 tttgcacact tcttccccca cctctaygcatcttcccccc actgcggtgt tcggcctgaa 900 ggtggtgggg gtgagggggg gtttggccattagcatttca tgtctttccc caaatgaaga 960 tgccctgcaa agggcagtna accacaaaaaaaaaaaaaaa aaaaacntgg ggggggggcc 1020 ccgttaacca ttttggcctn ataggggggnggtttttaaa aattaattgg gcccgg 1076 18 1379 DNA Homo sapiens SITE (639) nequals a,t,g, or c 18 ggcacgagca ccctcccaca cctccctgaa cttccatctgatcgacttca acttgctgat 60 ggtgaccacc atcgttctgg gccgccgctt cattgggtccatcgtgaagg aggcctctca 120 gagggggaag gtctccctct ttcgctccat cctgctgttcctcactcgct tcaccgttct 180 cacggcaaca ggctggagtc tgtgccgatc cctcatccacctcttcagga cctactcctt 240 cctgaacctc ctgttcctct gctatccgtt tgggatgtacattccgttcc tgcarctgaa 300 ttkcgamcty cgsaagacaa gcctcttcaa ccacatggcctccatggggc cccgggaggc 360 ggtcagtggc ctggcaaaga gccgggacta cctcctgacactgcgggaga cgtggaagca 420 gcacasaaga cagctgtatg gcccggacgc catgcccacccatgcctgct gcctgtcgcc 480 cagcctcatc cgcagtgagg tggagttcct caagatggacttcaactggc gcatgaagga 540 agtgctcgts agctccatgc tgagcgccta ctatgtggcctttgtgcctg tytggttcgt 600 gaagaacaca cattactatg acaagcgctg gtcctgtgnaactcttcctg ctggtgtcca 660 tcagcacctc cgtgatcctc atgcagcacc tgctgcntgccagctactgt gacctgctgc 720 acaaggccgc cgcccatctg ggctgttggc agaaggtggacccagcgctg tgctccaacg 780 tgctgcagca cccgtggact gaagaatgca tgtggccgcagggcgtgctg gtgaagcaca 840 gcaagaacgt ctacaaagcc gtaggccamw acaamgtggctatcccctct gacgtctccc 900 acttccgctt ccakttcttt ttcagcaaac ccctgcggatcctcaacatc ctcctgctgc 960 tggagggcgc tgtcattgtc tatcagctgt actccctaatgtcctctgaa aagtggcacc 1020 agaccatctc gctggccctc atcctcttca gcaactactatgccttcttc aagctgctcc 1080 gggaccgctt ggtattgggc aaggcctact catactctgctagcccccag agagacctgg 1140 accaccgttt ctcctgagcc ctggggtcac ctcagggacagcgtccaggc ttcagcaagg 1200 gctccctggc aaggggctgt tgggtagaag tggtggtgggggggacaaaa gacaaaaaaa 1260 tccaccagag ctttgtattt ttgttacgta ctgtttctttgataattgat gtgataagga 1320 aaaaagtcct atttttatac tcccaanmaa aaaaaaaaaanaaaaagcgg ccgaaagct 1379 19 1337 DNA Homo sapiens SITE (20) n equalsa,t,g, or c 19 ctggtgttgg gcctgagccn cctcaacaac tcctacaact tcagtttccacgtggtgatc 60 ggctctcagg cggaagaagg ccagtacagc ctgaacttcc acaactgcaacaattcagtg 120 ccaggaaagg agcatccatt cgacatcacg gtgatgatcc gggagaagaaccccgatggc 180 ttcctgtcgg cagcggagat gccccttttc aagctctaca tggtcatgtccgcctgcttc 240 ctggccgctg gcatcttctg ggtgtccatc ctctgcagga acacgtacagcgtcttcaag 300 atccactggc tcatggcggc cttggccttc accaagagca tctctctcctcttccacagc 360 atcaactact acttcatcaa cagccagggg ccaccccatc gaaggccttgccgkcatgta 420 ctacatcgca cacctgctga agggcgccct cctcttcatc accatcgccctgattggctc 480 aggctgggct tcatcaagta cgtcctgtcg gataaggaga agaaggtctttgggatcgtg 540 atccccatgc aggtcctggc caacgtggcc tacatcatca tcgagtcccgcgaggaaggc 600 gccacgaact acgtgctgtg gaaggagatt ttgttcctgg tggacctcatctgctgtggt 660 gccatcctgt tccccgtagt ctggtccatc cggcatctcc aggatgcgtctggcacagac 720 gggaaggtgg cagtgaacct ggccaagctg aagctgttcc ggcattactatgtcatggtc 780 atctgctacg tctacttcac ccgcatcatc gccatcctgc tgcaggtggctgtgcccttt 840 cagtggcagt ggctgtacma gctcttggtg garggctcca ccctggccttcttcgtgctc 900 acgggctaca agttccagcc cacagggaac aacccgtacc tgcagctgccccaggaggac 960 gaggaggatg ttcagatgga gcaagtaatg acggactctg ggttccgggaaggcctctcc 1020 aaagtcaaca aaacagccag cgggcgggaa ctgttatgat cacctccacatctcagacca 1080 aagggtcgtc ctcccccagc atttctcact cctgcccttc ttccacagcgtatgtgggga 1140 ggtggagggg tccatgtgga ccaggcgccc agctcccggg acsccggttcccggacaagc 1200 ccatttggaa gaagagtccc ttcctccccc caaatattgg gcagccctgtccttaccccg 1260 ggaccacccc tcccttccag ctatgtgtac aataatgacc aatctgtttggctaaaaaaa 1320 aaaaaaaaaa aactcga 1337 20 1390 DNA Homo sapiens SITE(1267) n equals a,t,g, or c 20 gccgttttgg ttcccggttg gtgcttcctgttcgcagctg cggcacttca aggttactga 60 ctttttatga tgtttggtgg ctatgagactatagawgcrt rsgrrgatga tytttatcga 120 gatgagtcat ctagtgaact gagtgttgatagtgaggtgg aatttcaact ctatagccaa 180 attcattatg cccaagatct tgatgatgtcatcagggagg aagagcatga agaaaagaac 240 tctgggaatt cggaatcttc gagtagtaaaccaaatcaga agaagctaat cgtcctttca 300 gatagtgagg tcatccagct gtcagatgggtcagaggtca tcactttgtc tgatgaagac 360 agtatttata gatgtaaagg aaagaatgttagagttcaag cacaagaaaa tgcccatggt 420 ctttcttctt ctcttcaatc taatgagctggttgataaga aatgcaagag tgatattgag 480 aagcctaaat ctgaagagag atcaggtgtaatccgagagg tcatgattat agaggtcagt 540 tcaagtgaag aggaagagag caccatttcagaaggtgata atgtggaaag ctggatgcta 600 ctgggatgtg aagtagatga taaagatgatgatatccttc tcaaccttgt gggatgtgaa 660 aactctgtta ctgaaggaga agatggtataaactggtcca tcagtgacaa agacattgag 720 gcccagatag ctaataaccg aacacctggaagatggaccc agcggtacta ttcagccaac 780 aaaaacatta tctgtagaaa ttgtgacaaacgtggtcatt tatcaaaaaa ctgcccctta 840 ccacgaaaag ttcgtcgctg cttcctgtgctccaggagag gacatctcct gtattcctgt 900 ccagcccccc tttgcgaata ctgtcctgtgcctaagatgt tggaccactc atgtcttttc 960 agacattcct gggataaaca gtgtgaccgatgtcatatgc taggccacta tacagatgct 1020 tgcacagaaa tctggaggca gtatcacctaacgaccaaac ctggaccacc caaaaagccg 1080 aagacccctt caagaccatc agccttagcatattgctatc actgcgcgca aaaaggccat 1140 tatggacacg aatgtccaga aagagaagtgtatgacccgt ctccagtatc tccattcatc 1200 tgctactatg rtgacaaata tgaaattcaggagagagaaa agagactaaa acaaaaaata 1260 aaagtantca agaaaaatgg ggttatcccagagccatcca agctacctta tataaaagca 1320 gcaaatgaga acccccacca tgatataaggaagggccgtg cctcatggaa aagcaacagg 1380 tggcctcaag 1390 21 1431 DNA Homosapiens 21 gcctgcagtc gacactagtg gatccaaaga attcggcctg tgcgagtaggcgcttgggca 60 ctcagtctcc ctggcgagcg acgggcagaa atctcgaacc agtggagcgcactcgtaacc 120 tggatcccag aaggtcgcga aggcagtacc gtttcctcag cggcggactgctgcagtaag 180 aatgtctttt ccacctcatt tgaatcgccc tcccatggga atcccagcactcccaccagg 240 gatcccaccc ccgcagtttc caggatttcc tccacctgta cctccagggaccccaatgat 300 tcctgtacca atgagcatta tggctcctgc tccaactgtc ttagtacccactgtgtctat 360 ggttggaaag catttgggcg caagaaagga tcatccaggc ttaaaggctaaagaaaatga 420 tgaaaattgt ggtcctacta ccactgtttt tgttggcaac atttccgagaaagcttcaga 480 catgcttata agacaactct tagctaaatg tggtttggtt ttgagctggaagagagtaca 540 aggtgcttcc ggaaagcttc aagccttcgg attctgtgag tacaaggagccagaatctac 600 cctccgtgca ctcagattat tacatgacct gcaaattgga gagaaaaagctactcgttaa 660 agttgatgca aagacaaagg cacagctgga tgaatggaaa gcaaagaagaaagcttctaa 720 tgggaatgca aggccagaaa ctgtcactaa tgacgatgaa gaagccttggatgaagaaac 780 aaagaggaga gatcagatga ttaaaggggc tattgaagtt ttaattcgtgaatactccag 840 tgagctaaat gccccctcac aggaatctga ttctcacccc aggaagaagaagaaggaaaa 900 gaaggaggac attttccgca gatttccagt ggccccactg atcccttatccactcatcac 960 taaggaggat ataaatgcta tagaaatgga agaagacaaa agagacctgatatctcgaga 1020 gatcagcaaa ttcagagaca cacataagaa actggaagaa gagaaaggcaaaaaggaaaa 1080 agaaagacag gaaattgaga aagaacggag agaaagagag agggagcgtgaaagggaacg 1140 agaaaggcga gaacgggaac gagaaaggga aagagaacgt gaacgagaaaaggagaaaga 1200 acgggagcgg gaacgagaac gggataggga ccgtgaccgg acaaaagagagagaccgaga 1260 tcgggatcga gagagagatc gtgaccggga tagagaaagg agctcagatcgtaataagga 1320 tcgcattcga tcaagagaaa aaagcagaga tcgtgaaagg gaacgagagcgggaaagaga 1380 gagagagaga gaacgagagc gagaacgaga acgggagcga gagagagaag c1431 22 2539 DNA Homo sapiens SITE (1283) n equals a,t,g, or c 22gggtgcagga gtgccacccc cagggccctg tcaacctctc ttttctcctc catggctgtc 60tgcctgcgta tctgtctctg agaatcctcg gggcggtcag gggatgtcag gaggggaagg 120agccgccctc cctatcttgc tgctcctctt ggcactcagg ggcaccttcc atggagccag 180accgggtgga ggggcttctg ggatttggtg tctgctgctg ccagagcagg aacccccagt 240ctaggacttg ggcattttaa cagggagaaa gtagtggctt cccttttctc tctctcctcc 300tttttccctt taagcccaca gattcaggtc atgccaaaag ctctctggtt gtaacctgga 360gacatgtgga ggggaatggc gatgggatta taggactctc cccatctcgg gccctgaccc 420tgacccttgc caccaaccca aagacagctg gtgggtttcc ccttggagam aatcctgcgt 480ttgcctgggc cggccctggc tgccctcagc tttcgctgat ctgcccggcc tggagcctcc 540catcaccccg cttcttgttg ggcctcaggc actggttacc agaagggggt ctgggtctgc 600tcaggaatca tgttttgtag cacctcctgt tggaggggtg gagggatgtt cccctgagcc 660aggctgagac tagaacccca tcttccctga gccaggctga gactagaacc ccatcttccc 720caccacgcca cccctgtgst kgctacagga gcacagtagt gaaggcctga gctccaggtt 780tgaaagaccc aactggagcg tggggcgggc aggcaggggt tagtgaaagg acacttccag 840ggttaggaca gagcatttag ccttctggaa gaacccctgc ctggggtggg actgtgcagg 900ccagagaagg tggcatgggc ctgaacccac ctggactgac ttctgcactg aagccacaga 960tggagggtag gctggtgggt gggggtggtt cgttctctag ccggggcaga cacccagctg 1020gctgggtcct tcctcagcct tgcctcctcc tgtccccaac cctttccttt cctcctgctt 1080gcggactgct ggtcccctct ccttccctcc ttccagctgt ttctagttac cacctacccc 1140tgggccgtgg actgatcaga ccagcattca aaataaaagt ttgttccaag ttgacagtgt 1200ggtgctccct gcccagcccc tccaggtgga ggtgctgcca cgggaacgca gttgctctgc 1260ctgccctggg cccctggcga cantgggagc agggcagtgc tgtgaggagc ccagctttcc 1320cagtcaggca ggcatggctt ccgtgttcag gctccctcac cagctggtga cacgggacaa 1380gcttacaaac cttctctgaa cctcagtttt ctcatttaca agaggcaaag catccatcac 1440cttgtgtgga ttcaragaat gtraggccct ggggtgtcct acacaaggga aaggcttgct 1500cagtgagcgg tctgcacacc gttagccacc ctgccacctc tgtgccctgg gcaggctcca 1560aaggaaagct ctggctggga ctgccrggag tctcacacgc tcctgttgac attcccagca 1620gcygcccctg aggtcgatgt ttgttctgtt tttctttttc ttttttgaga cggagtctcg 1680ctgtgttgcc aggctggagt gcagtggtgt gatctctgct cactgcaacc tccgcctgcc 1740agtttcaagt gattctctgc ctcagccttc tgagtagctg ggactacagg tgcacgccac 1800cacgcccagc taactttttg tatttwagta gagacagggt ttcgccatgt cggccagggt 1860ggtcttgatc tcctgacctc atgatccacc cgcctcagcc tcccaaagtg ctgggattac 1920aggtatgagc caccgcaccg ggcctgttct atttttctag ttaagggaac tgaagctcag 1980araggtgtca ccagcargtg ttcattccca tgccagcctt gccccccggc ttttcccagg 2040caggctcctg cgtgcccact ggctccagcc tggtcctctg tctcttggct gcttcactcc 2100tgctctttgt cccgactctg gccctgctta caggggccac tacctgctgg tgcctccata 2160acaagcgtct ggcgttgaga cccctggcat ggcaggggct ttggggtctg gtttccacaa 2220ggcttagcca tggcagaacc tcgttttatt ttaactcttt gcccctacaa acaaacagca 2280gtacttgcca gaaccattct tgggattcag gagctcgggc gactgccttg gcctctggcc 2340gcacccagga gggtggggtt ggatctgtgt agttgccagg cccacacctg ccagcagggg 2400gctgactgga tccatgcttt actgtgttta atgggggtaa caggggtccc tacagccctc 2460ccagytaaam atttggaaca aaacaccagc ccttttgtag tggatgcaga ataaaattgt 2520taatccaatc aaaaaaaaa 2539 23 1041 DNA Homo sapiens 23 tcgacccacgcgtccgccca cgcgtccgcc cacgcgtccg ggcgcaggac gtgcactatg 60 gctcggggctcgctgcgccg gttgctgcgg ctcctcgtgc tggggctctg gctggcgttg 120 ctgcgctccgtggccgggga gcaagcgcca ggcaccgccc cctgctcccg cggcagctcc 180 tggagcgcggacctggacaa gtgcatggac tgcgcgtctt gcagggcgcg accgcacagc 240 gacttctgcctgggctgcgc tgcagcacct cctgccccct tccggctgct ttggcccatc 300 cttgggggcgctctgagcct gaccttcgtg ctggggctgc tttctggctt tttggtctgg 360 agacgatgccgcagagagag aagttcacca cccccataga ggagaccggc ggagagggct 420 gcccagctgtggcgctgatc cagtgacaat gtgccccctg ccagccgggg ctcgcccact 480 catcattcattcatccattc tagagccagt ctctgcctcc cagacgcggc gggagcaagc 540 tcctccaaccacaagggggg tggggggcgg tgaatcacct cygaggcctg ggcccagggt 600 tcaggggaacttccaaggtg tctggttgcc ctgcctctgg ctccagaaca gaaagggagc 660 ctcacgctggctcacacaaa acagctgaca ctgactaagg aactgcagca tttgcacagg 720 ggaggggggtgccctccttc ctagaggccc tgggggccag gctgacttgg ggggcagact 780 tgacactaggccccactcac tcagatgtcc tgaaattcca ccacgggggt caccctgggg 840 ggttagggacctatttttaa cactaggggg ctggcccact aggagggctg gccctaagat 900 acagacccccccaactcccc aaagcgggga ggagatattt attttgggga gagtttggag 960 gggagggagaatttattaat aaaagaatct ttaactttaa aaaaaaaaaa aaaaaagggc 1020 ggccgctctagaggatccct c 1041 24 1962 DNA Homo sapiens SITE (452) n equals a,t,g, orc 24 acccacgcgt ccggtacaaa acacagtttt attctatgaa aattttgaga ttattagaaa60 cattagattt agggttgcat attaaaaact atatccattt tgccttatta tttagtgtct 120cactcaggat ataacacact ataatagaaa atgtagactt cagaatcagg tatatttgag 180atggtttgta tactggttct gacacttgtt agctattcat ctttggtaaa ttccccatta 240ccctttgtkc acctatwtgt ggggatcagt gcatagtgtg tgtwaagcat ttaatacctg 300gcaagtgttc agcaaatttt ttgttctata tatttattat ttgattattg gccctgagga 360gtaggtgttt gtttgtttgt ttgtttgttt agttttattt ctcatctcct caggaacaca 420aatgaaactt ggatattgtt atggtgcttt tnataatata tttattattt tcagcaattn 480attcttgtta aaacaatttc ttatgacaag ttactcatct tcaatggtga gaagaaatct 540agctcagaat aatatatttt tagtgtttgt atctctggat actcattttg ctcattgcca 600cgtaaagtaa aaaaatacat aaattagctt attccaatgt aatatcttca ggatagtcat 660gggcaaggaa ttaatcacat taagagataa ctgcaactaa gcactatttg aggtgacttc 720tgtggaaaaa aaattaatyc tttaccattg cagcgttctg ccctaggtcc aaatgttacc 780aaaatcactc tagaatcttt tcttgcctgg aagaaaagga aaagacaaga aaagattgat 840aaacttgaac aagatatgga aagaaggaaa gctgacttca aagcagggaa agcactagtg 900atcagtggtc gtgaagtgtt tgaatttcgt cctgaactgg tcaatgatga tgatgaggaa 960gcagatgata cccgctacac ccagggaaca ggtggtgatg aggttgatga ttcagtgagt 1020gtaaatgaca tagatttaag cctgtacatc ccaagagatg tagatgaaac aggtattact 1080gtagccagtc ttgaaagatt cagcacatat acttcagata aagatgaaaa caaattaagt 1140gaagcttctg gaggtagggc tgaaaatggt gaaagaagtg acttggaaga ggacaacgag 1200agggagggaa cggaaaatgg agccattgat gctgttcctg ttgatgaaaa tcttttcact 1260ggagaggatt tggatgaact agaagaagaa ttaaatacac ttgatttaga agaatgacac 1320caaacacatc gctgaaaaaa ttaagtcagc tcagcacgag ttgaaattga ctacattaat 1380ttctttccac ctagaatcaa caggatgttt atttcctatg ctgattctgg aggagttaac 1440ctcctgcaaa aaaggcatct tgtccctaca tcttctcttc tgactttggc tacatctcat 1500agtaagttca gagtagttca tgataaattg aaaatataat ggtcattgca gaaaatgatt 1560gatgttgtaa ctgtccaccc aagtaagaag tgtatctgcc tttccatctt ttggttttca 1620tttgggcatg tgctattacc agaaacaaca aacttatatt taaaataccc ttcatttgac 1680acagttttta atgagtgatt taatttcctc tgtatttgta tgtttagaag actgcctaaa 1740acatgagcac tgtacttcat aaaggaaacg cgtatgcaga ttcagtattg tgtatctttg 1800gacaattaga tggacattta aaatggaact tcttttatct gacaggatca gctacaatgc 1860cctgtgttaa attgtttaaa agtttccctt ttcttttttg ccaataaagt tgtaaataaa 1920gaccatcata cattaaaatc caaaaaaaaa aaaaaaaaaa aa 1962 25 1228 DNA Homosapiens SITE (580) n equals a,t,g, or c 25 ggctgcccag gccccgcactggaagagcct ccagcagcaa gatgtgaccg ytgtgccgat 60 gagccccagc agccactccccagaggggag gcctccacct ctgctgcctg ggggtccagt 120 gtgtaaggca gctgcatctgcaccgagctc cctcctggac cagccgtgcc tctgccccgc 180 accctctgtc cgcaccgctgttgccctgac aacgccggat atcacattgg ttctgccccc 240 tgacatcatc caacaggaagcgtcaccctg agggaggaga cagaagcctg ggccaggtga 300 acagtggtat agcagccactccagcctctg ctgcagcagc caccctggat gtggctgttc 360 ggagaggcct gtcccacggagcccagaggc tgctgtgcgt ggccctggga cagctggacc 420 ggcctccaga cctcgcccatgacgggagga gtctgtggct gaacatcagg ggcaaggagg 480 cggctgccct atccatgttccatgtctcca cgccactgcc agtgatgacc ggtggtttcc 540 tgagctgcat cttgggcttggtgctgcccc tggcctatgn ttccagcctg acctggtgct 600 ggtggcgctg gggcctgccantgcctgcag ggcccccacg ctgcactcct ggctgcaatg 660 cttcgggggc tggcagggggccgagtcctg gccctcctgg aggagaactc cacaccccag 720 ctagcaggga tcctggcccgggtgctgaat ggagaggcac ctcctagcct aggcccttcc 780 tctgtggcct ccccagaggacgtccaggcc ctgatgtacc tgagagggca gctggagcct 840 cagtggaaga tgttgcagtgccatcctcac ctggtggctt gaaatcggcc aaggtgggag 900 catttacacc gcagaaatgacaccgcacgc cagcgccccg cggccgcgat ccggacccca 960 agcccacggc tccctcgactctggggcacg gaaccccgcc cactcccaat ccccgcgccc 1020 cgccctctcc cacccgtgcttcccccgctc cacccctcac ctcacctcgc cccsgcccca 1080 cccatcgcgc cccggcccgtcccatcgagg cccatgcaac ccacgctcgg tyccgttccg 1140 gcccctgcgc tckcgctknsttcgctcccc gcccttgcgc cgttagtaaa catcgctcaa 1200 acgaaaaaaa aaaaaaaaaaaaactcga 1228 26 1340 DNA Homo sapiens SITE (847) n equals a,t,g, or c26 aattcggcag agagatggcc gcccccgtgg atctagagct gaagaaggcc ttcacagagc 60ttcaagccaa agttattgac actcaacaga aggtgaagct cgcagacata cagattgaac 120agctaaacag aacgaaaaag catgcacatc ttacagatac agagatcatg actttggtag 180atgagactaa catgtatgaa ggtgtaggaa gaatgtttat tcttcagtcc aaggaagcaa 240ttcacagtca gctgttagag aagcagaaaa tagcagaaga aaaaattaaa gaactagaac 300agaaaaagtc ctacctggag cgacgttaaa ggaagctgag gacaacatcc gggagatgct 360gatggcacga agggcccagt agggagcctc tctgggaagc tcttcctcct gcccctccca 420ttcctggtgg gggcagagga gtgtctgcag ggaaacagct tctcctctgc cccgatggat 480gctttatttg gatggcctgg caacatcaca ttttctgcat caccctgagc cccatttgct 540tcccagccct ggagttttta cccggctttg ctgccacctc tgcccaggac ackcttccct 600ctcgggatgt gtgatgaact cccaggagag ggaagatggg agccagggca agataggaag 660ctctgcctga gctttccact aggcacgcca gccagaccaa taaaaagcgt ctgtcccact 720ctgctaagcc tggttttctt gagcagaggg atggaacaga gggtgagaga ggcagtggcc 780gtctccacct cagctcctgc tccctctgca tcagagccct tcctttcttg ggggatgggc 840cttgccntct tctcttttcc cttcctgtac ctttgactaa cgctcagctt ccgggcctgc 900atgcagtaga cagaagagga agaaagaaca gatgttcaca gctgaatctc agtgaacaga 960atagcagtcc ctggatggca gtctgcctaa agattccttt ccctgccttc tcccatacat 1020tccaaaagga agttcaacag taagcagcac ctccaagact gtctccttty ggccartatc 1080ataagatgga cgccataatc ctgaggcctc ctagaggctg agggggcaac ggtgtgatcc 1140agctggctca tcccagccag gtgggccaat tattcaattt tcaagaattt tgttgcaagc 1200cagttgtcaa acacagccat tataattatg taaatttgca aattatgtta aaaacaagga 1260caataaatat tcaaaatgca tccctaawwa aaaaaaaaaa aangggnggc cgcnctaggg 1320gatccaagct tacgtacgcg 1340 27 806 DNA Homo sapiens 27 accttcttccatgtttagtc ccttgggctc tgctaccctc ctgctggagg tgagagcatc 60 ctgtgtgcaaccagagatgc cctctggctt tcagacctgc ctgcttttca ccctcagccc 120 tttctcactcagcaaaattg tgggggtccc tagtcagcag ctccctgggc agctctctga 180 gcaaggtggtctctgtggtc atgaaggaga gccggctagg acagtgccgg aaactcagct 240 gcctctccccttcaactcag ctggcccccc gcacctgaag tgcacaggag ccgggaagag 300 agtctggagcccaccccgga gggcagcaca ggaggtgtct ctgcagctgg tgtcctgcca 360 cccctgcaggcagcacacgt cccgggcatt ctccttagcc acagacagaa cagccagtgc 420 cagagtctgctgtcgttccc ctttaagcac actcattcac cacacccgag gaggccagag 480 gtgcagggagcatgggctgt cgcttcccct ttaagcacac tcattcacca cacccgagga 540 ggccagaagtgcagggagca tgggctgggt gcacctccgc aggagagaag gctgagccac 600 cgccgtcccgggagcccggc tcccaggcct ctcgttttcc cctacctccc taagactttt 660 ctgtcactctctggccattg aaaggcttct gttccttaaa gtgctgttac actctccttt 720 cccaggatgcagcaagccaa aacagtacca ctgcacgtca gcctgggtga cagagtgaga 780 ccctatcttaaaaaaaaaaa aaaaaa 806 28 696 DNA Homo sapiens SITE (9) n equals a,t,g,or c 28 gagttcccna cgcggtggcg nccgttttag aaattagtgg atccccccgggctggcaggg 60 aattcggcac gagcacagag gaaagcgggt gcccggcatg gccatcctgatgttgctggc 120 gggatcccca tgcaccttgt ccttctccac tgatactggc agctcggctcctggacccaa 180 gatcccttga gtggaattct gcagtgcaag agcccttcgt gggagctgtcccatgtttcc 240 atggtcccca gtctcccctc cacttggtgg ggtcaccaac tactcaccagaagggggctt 300 accaagaaag ccctaaaaag ctgttgactt atctgcgctt gttccaactcttatgccccc 360 aacctgccct accaccacca cgcgctcagc ctgatgtgtt tacatggtactgtatgtatg 420 ggagagcaga ctgcaccctc cagcaacaac agatgaaagc cagtgagcctactaaccgtg 480 ccatcttgca aactacactt taaaaaaaac tcattgcttt gtattgtagtaaccaatatg 540 tgcagtatac gttgaatgta tatgaacata ctttcctatt tctgttctttgaaaatgtca 600 gaaatatttt tttctttctc attttatgtt gaactaaaaa ggattaaaaaaaaaatctcc 660 agamaaaaaa aaaaaaaaaa aaattactgc ggtccg 696 29 1007 DNAHomo sapiens SITE (922) n equals a,t,g, or c 29 aattcggcac gaggaaaaaataccatttgt gtatgatacc caatttggat ctcaatttgg 60 atagagattt ggtgcttccagatgtragtt atcaggtgga atccagtgag gaggatcagt 120 ctcagactat ggatcctcaaggacaaactc tgctgctttt tctctttgtg gatttccaca 180 gtgcatttcc agtccagcaaatggaaatct ggggagtcta tactttgctc acaactcatc 240 tcaatgccat ccttgtggagagccacagtg tagtgcaagg ttccatccaa ttcactgtgg 300 acaaggtctt ggagcaacatcaccaggctg ccaaggctca gcagaaacta caggcctcac 360 tctcagtggc tgtgaactccatcatgagta ttctgactgg aagcactagg agcagcttcc 420 gaaagatgtg tctccagacccttcaagcag ctgacacaca agagttcagg accaaactgc 480 acaaagtatt tcgtgagatcacccaacacc aatttcttca ccactgctca tgtgaggtga 540 agcagctaac cctagaaaaaaaggactcag cccagggcac tgaggacgca cctgataaca 600 gcagcctgga gctcctagcagataccagcg ggcaagcaga aaacaagagg ctcaagaggg 660 gcagcccccg catagaggagatgcgagctc tgcgctctgc cagggccccg agcccgtcag 720 aggccgcccc gcgccgcccggaagccaccg cggcccccct cactcctaga ggaagggagc 780 accgcgaggc tcacggcagggccctggcgc cgggcagggc gagcctcgga agccgcctgg 840 aggacgtgct gtggctgcaggaggtctcca acctgtcaga gtggctgagt cccagccctg 900 ggccctgagc cgggtccccttncgcaagcg cccaccgatc cggargctgc gggcagccgt 960 tatcccgtgg tttaataaagtgccgcgcgc tcaccaaaaa aaaaaaa 1007 30 2026 DNA Homo sapiens 30gaattcggca cgagcacgga tccgttgcgg ctgcagctct gcagtcgggc cgttccttcg 60ccgccgccag gggtagcggt gtagctgcgc agcgtcgcgc gcgctaccgc acccaggttc 120ggcccgtagg cgtctggcag cccggcgcca tcttcatcga gcgccatggc cgcagcctgc 180gggccgggag cggccgggta ctgcttgctc ctcggcttgc atttgtttct gctgaccgcg 240ggccctgccc tgggctggaa cgaccctgac agaatgttgc tgcgggatgt aaaagctctt 300accctccact atgaccgcta taccacctcc cgcaggctgg atcccatccc acagttgaaa 360tgtgttggag gcacagctgg ttgtgattct tataccccaa aagtcataca gtgtcagaac 420aaaggctggg atgggtatga tgtacagtgg gaatgtaaga cggacttaga tattgcatac 480aaatttggaa aaactgtggt gagctgtgaa ggctatgagt cctctgaaga ccagtatgta 540ctaagaggtt cttgtggctt ggagtataat ttagattata cagaacttgg cctgcagaaa 600ctgaaggagt ctggaaagca gcacggcttt gcctctttct ctgattatta ttataagtgg 660tcctcggcgg attcctgtaa catgagtgga ttgattacca tcgtggtact ccttgggatc 720gcctttgtag tctataagct gttcctgagt gacgggcagt attctcctcc accgtactct 780gagtatcctc cattttccca ccgttaccag agattcacca actcagcagg acctcctccc 840ccaggcttta agtctgagtt cacaggacca cagaatactg gccatggtgc aacttctggt 900tttggcagtg cttttacagg acaacaagga tatgaaaatt caggaccagg gttctggaca 960ggcttgggaa ctggtggaat actaggatat ttgtttggca gcaatagagc ggcaacaccc 1020ttctcagact cgtggtacta cccgtcctat cctccctcct accctggcac gtggaatagg 1080gcttactcac cccttcatgg aggctcgggc agctattcgg tatgttcaaa ctcagacacg 1140aaaaccagaa ctgcatcagg atatggtggt accaggagac gataaagtag aaagttggag 1200tcaaacactg gatgcagaaa ttttggattt ttcatcactt tctctttaga aaaaaagtac 1260tacctgttaa caattgggaa aaggggatat tcaaaagttc tgtggtgtta tgtccagtgt 1320agctttttgt attctattat ttgaggctaa aagttgatgt gtgacaaaat acttatgtgt 1380tgtatgtcag tgtaacatgc agatgtatat tgcagttttt gaaagtgatc attactgtgg 1440aatgctaaaa atacattaat ttctaaaacc tgtgatgccc taagaagcat taagaatgaa 1500ggtgttgtac taatagaaac taagtacaga aaatttcagt tttaggtggt tgtagctgat 1560gagttattac ctcatagaga ctataatatt ctatttggta ttatattatt tgatgtttgc 1620tgttcttcaa acatttaaat caagctttgg actaattatg ctaatttgtg agttctgatc 1680acttttgagc tctgaagctt tgaatcattc agtggtggag atggccttct ggtaactgaa 1740tattaccttc tgtaggaaaa ggtggaaaat aagcatctag aaggttgttg tgaatgactc 1800tgtgctggca aaaatgcttg aaacctctat atttctttcg ttcataagag gtaaaggtca 1860aatttttcaa caaaagtctt ttaataacaa aagcatgcag ttctctgtga aatctcaaat 1920attgttgtaa tagtctgttt caatcttaaa aagaatcaat aaaaacaaac aaggggaaaa 1980aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa actcga 2026 31 699 DNA Homosapiens SITE (2) n equals a,t,g, or c 31 gngttttttc cagccaggaagtgaccgnta ctgcagcacg aganagattg gttgggttgg 60 ttgraaatga cyctgaacatttatttccat tgcaatttct gtggctgagg agacttaaac 120 tttacaagta ttatccttttaagatcattt taattttagt tgagtgcaga gggcttttat 180 aacaaacgtg cagaaattttggagggctgt gatttttcca gtattaaaca tgcatgcatt 240 aatcttgcag tttattttctcattgtgtat gtatatatcg cttttctctg cagcacgatt 300 tctcttttga taawkccctttagggcacaa ctagttatca gtaactgaat gtatcttaat 360 cattatggct gcttctgttttttcattaac aaaggttatt catatgttag catatagttt 420 ctttgcaccc actatttatgtctgaatcat ttgtcacaag agagtgtgtg ctgatgagat 480 tgtaagtttg tgtgtttaaacttttttttg agcgagggaa gaaaaagctg tatgcatttc 540 attgctgtct acaggtttctttcagattat gttcatgggt ttgtgtgtat acaatatgaa 600 gaatgatctg aagtaattgtgctgtattta tgtttattca ccagtctttg attaaataaa 660 aaggaaaacc agaaaaaaaaaaaaaaaaaa aaaaaaaaa 699 32 1264 DNA Homo sapiens SITE (1057) n equalsa,t,g, or c 32 ggcacgaggg cactgtttcc tcagtccatg gctgagtaca tcaccggtgttttctctctt 60 attcctccca tcaagcctaa aaggaatctc tattggagat actgccattagtgttccttt 120 tataggtgag gaactgaggc atakagggtt ccccagttga accaactgataaatagtaga 180 acttggattt taattcagtc ttgatgccag ggataaggct cttactttctaccttaggct 240 atttctagga aacgcaggag agtgttgaag gggcagagaa agggatccagttcctttctg 300 tcccgcatcc tagtccctga gaagcaaaga araatgtgtg gcttcttttgctttgctttt 360 gttgtcatcc cacacatctc caggggamct gggctcttga tcttggsctcttccccttta 420 actgttaagt gggagcargt aagggggtac agtagggctg gcctggagttagaggcttgg 480 atgccttagc tcctctgtct gcactccaga actgcctgac ttcatttcgtatgttgtcct 540 ttgttttgac aattgatcca tgtcccagtc cgtctcttct tccttcttgatacttacact 600 gcttctttct gttggtttcc agtgtttaac actgtataca acagtgacgacaacgtgttt 660 gtgggggccc ccacgggcag cgggaagact atttgtgcag agtttgccatcctgcgaatg 720 ctgctgcaga gctcggaggg gcgctgtgys twcwtcaccm ccatggaggccctggccaga 780 rcaggtatga cgtggcgctg tgtcatgtga atttcccaag aagcatttcatctgtgattc 840 cgtatgaagg ctttctaagc cctgaaattt gcagggtcat ttcctcagtttgtgtattaa 900 agaaaagctg ccccagccaa gcgtggtggc tcacgcctgt aatcccagcactttgggagg 960 ccgaggcggg cagatctccg gagatcagga gttcgagacc agcctggccaacatggtgra 1020 accctgtctc tactaaaawt acagaaatta gctgggngtg gtggtgtgcgcctgtaatcc 1080 cagctacttg gaaggctgag gcaggagaat cgcttgaacc cgggaggcggaggttgcagt 1140 gagccaagtt cgcaccactg cactccagcc tgggcaacaa gagcgagacttcatctcaaa 1200 aaaaaaaaaa aaaaactcga gggggggccc ggtacccaat tcgccctatagtgatcgtat 1260 taca 1264 33 997 DNA Homo sapiens SITE (855) n equalsa,t,g, or c 33 attggaagtt gttttgcaac ctgggctttt atacagaaga atacgaatcacaggtgtgtg 60 agcatctact taattaattt gcttacagcc gatttcctgc ttactctggcattaccagtg 120 aaaattgttg ttgacttggg tgtggcacct tggaagctga agatattccactgccaagta 180 acagcctgcc tcatctatat caatatgtat ttatcaatta tcttcttagcatttgtcagc 240 attgaccgct gtcttcagct gacacacagc tgcaagatct accgaatacaagaacccgga 300 tttgccaaaa tgatatcaac cgttgtgtgg ctaatggtcc ttcttataatggtgccaaat 360 atgatgattc ccatcaaaga catcaaggaa aagtcaaatg tgggttgtatggagtttaaa 420 aaggaatttg gaagaaattg gcatttgctg acaaatttca tatgtgtagcaatattttta 480 aatttctcag ccatcatttt aatatccaat tgccttgtaa ttcgacagctctacagaaac 540 aaagataatg aaaattaccc aaatgtgaaa aaggctctca tcaacatacttttagtgacc 600 acgggctaca tcatatgctt tgttccttac cacattgtcc gaatcccgtataccctcagc 660 cagacagaag tcataactga ttgctcaacc aggatttcac tcttcaaagccaaagaggct 720 acactgctcc tggctgtgtc gaacctgtgc tttgatccta tcctgtactatcacctctca 780 aaagcattcc gctcaaaggt cactgagact tttgcctcmc ctaaagagaccaaggtyaga 840 aagaaaaatt aagangtgga aataatggct aaaagacagg ntttttgtggtaccaattct 900 gggctttatg ggaccntaaa gttattatag cttggaaggt aaaaaaaaaaaaagggnggg 960 cgctctagag gttccccgag gggccagctt agggtgc 997 34 1914 DNAHomo sapiens SITE (1889) n equals a,t,g, or c 34 gtgtgagagg cctctctggaagttgtcccg ggtgttcgcc gctggagccc gggtcgagag 60 gacgaggtgc cgctgcctggagaatcctcc gctgccgtcg gctcccggag cccagccctt 120 tcctaaccca acccaacctagcccagtccc agccgccagc gcctgtccct gtcacggacc 180 ccagcgttac catgcatcctgccgtcttcc tatccttacc cgacctcaga tgctcccttc 240 tgctcctggt aacttgggtttttactcctg taacaactga aataacaagt cttgatacag 300 agaatataga tgaaattttaaacaatgctg atgttgcttt agtaaatttt tatgctgact 360 ggtgtcgttt cagtcagatgttgcatccaa tttttgagga agcttccgat gtcattaagg 420 aagaatttcc aaatgaaaatcaagtagtgt ttgccagagt tgattgtgat cagcactctg 480 acatagccca gagatacaggataagcaaat acccaaccct caaattgttt cgtaatggga 540 tgatgatgaa gagagaatacaggggtcagc gatcagtgaa agcattggca gattacatca 600 ggcaacaaaa aagtgaccccattcaagaaa ttcgggactt agcagaaatc accactcttg 660 atcgcagcaa aagaaatatcattggatatt ttgagcaaaa ggactcggac aactatagag 720 tttttgaacg agtagcgaatattttgcatg atgactgtgc ctttctttct gcatttgggg 780 atgtttcaaa accggaaagatatagtggcg acaacataat ctacaaacca ccagggcatt 840 ctgctccgga tatggtgtacttgggagcta tgacaaattt tgatgtgact tacaattgga 900 ttcaagataa atgtgttcctcttgtccgag aaataacatt tgaaaatgga gaggaattga 960 cagaagaagg actgccttttctcatactct ttcacatgaa agaagataca gaaagtttag 1020 aaatattcca gaatgaagtagctcggcaat taataagtga aaaaggtaca ataaactttt 1080 tacatgccga ttgtgacaaatttagacatc ctcttctgca catacagaaa actccagcag 1140 attgtcctgt aatcgctattgacagcttta ggcatatgta tgtgtttgga gacttcaaag 1200 atgtattaat tcctggaaaactcaagcaat tcgtatttga cttacattct ggaaaactgc 1260 acagagaatt ccatcatggacctgacccaa ctgatacagc cccaggagag caagcccaag 1320 atgtagcaag cagtccacctgagagctcct tccagaaact agcacccagt gaatataggt 1380 atactctatt gagggatcgagatgagcttt aaaaacttga aaaacagttt gtaagccttt 1440 caacagcagc atcaacctacgtggtggaaa tagtaaacct atattttcat aattctatgt 1500 gtatttttat tttgaataaacagaaagaaa ttttgggttt ttaatttttt tctccccgac 1560 tcaaaatgca ttgtcatttaatatagtagc ctcttaaaaa aaaaaaaaac ctgctaggat 1620 ttaaaaataa aaatcagaggcctatctcca ctttaaatct gtcctgtaaa agttttataa 1680 atcaaatgaa aggtgacattgccagaaact taccattaac ttgcactact agggtaggga 1740 ggacttaggg atgtttcctgtgtcgtatgt gcttttcttt ctttcatatg atcaattctg 1800 ttggtatttt cagtatctcatttctcaaag ctaaagagat atacattctg gatacttggg 1860 aggggaataa attaaagttttcacactgna aaaaaaaaaa aaaaaaaaac tcga 1914 35 1020 DNA Homo sapiens SITE(18) n equals a,t,g, or c 35 gtataattat aaatttgntc ggttcnaccg gtcctgtgttgcytaaaaac accttataaa 60 agaggagagt atttgataag caattttcat agtagtaaagttttttttca tctcttaaac 120 taaattgacc atgcatataa tattctttgt ttaaatgaaagcatactgtt gaaacccgca 180 gtgttgcatt tagaaaacag ttgaacagaa tgtcaatgtgcattcatgca aaaaaacatt 240 taatctgcat ctgttttaga aaagggggaa atgaagcaacttgtctaaaa atactgcttt 300 acaaagcatt tcagcctttc cccctcagtt ttgcattgattttttgacaa gtctgtagag 360 cctaatagtt tccatcaaag gcctagatct cttatttagcatttttttca gctcttctct 420 cagaagttca gctgttgaaa cgaaaactgt actttgtaccctcacataca aagggatcaa 480 atttgacctg gtgttatttt agccccaaat ttatgacattacacaatatt aaaatgtaaa 540 tgtttcttta cccaaactac ttctagatat tctagtatttgcttctggtg gaattaaatg 600 acggtaaaat tggctaatta tttgaatgaa tgaatggatggatgttttgc atgctcaatt 660 tctaggtcct ttgtctagaa aggaaatttg cctcagttgaattagtgaaa tatttctgtc 720 gttgatatta aaagtgactt ctgagtacag ttaagttcctcctatttgcc actgggctgt 780 tggttagaag cataggtaac tgattaagta ggtatgatactgcatttgaa ataagtggac 840 acaaactatc ctttctccac catggactca atctgagaacaacagcattc atttccattc 900 atttccatac tggcttttga ttatatgcag attcctagtagcatgcctta cctacagcac 960 tatgtgcatt tgctgtcaca ataaagtata ttttgtcttgcaaaaaaaaa aaannaangg 1020 36 781 DNA Homo sapiens 36 aactcctgacctcaagtgct ccacctgcgt tggcttccca aagtgctggg atacaggagt 60 ragccactgcgcctggctga tcccagcact tttmaaatga tgccgctcaa agccgtgact 120 tggcctactttgaacagcaa acttgttgct gctgttgtca acctgaaggc ctctcaaatg 180 ccagcttcaagcagggtgtg aattggccag tgtcagatct caggagtcct gtgttgagag 240 tgtggctttcagctgcgggg agctgcactt ggtggggaaa gccaggcagg tcaccctcac 300 agccagataatgtggaggtc agaacccaag gaagggagtg agacctccac tcccagtggg 360 ggacctggccacccatcctt ggggacctga gaaagcgtac ttcaccttgg ggtgaaggct 420 gggtggggccagagggacca gtgccctcct cagtgcttag gggcagagcc acctgcagca 480 atggtatctgcatattagcc cctctccacc ttctttctcc cgctgaatca tttccctcaa 540 agcccaagagctgtcactgc ttctttctcc ctgggaagaa tgcgtggact ctgcctggtg 600 atagactgaagccagaacag tgccacaccc tcgccttaat tccttgctag gtgttctcag 660 atttatgagacttcttagtc aaatatgagg gaggttggat gtggtggctt gtgcctgtaa 720 tcccagcattttgggaagcc gaggtgggag gatcccttga agccaggagt ttgagacaag 780 c 781 37 966DNA Homo sapiens SITE (8) n equals a,t,g, or c 37 ccactatngg caattggtaccggccccccc tcgaggaaga taaggtgcag ttcatgccag 60 aatccccagc ttcccatgcaggctggggac atagtgggtt ctcccgaaat actgggtcac 120 ttgaatttga tatgatgtatatatattcac ctctagtcca taggtacata tagtctatat 180 attaaaaaga cattggattttgacttaaac tagatgtttc tcaagcacac caagacggtg 240 ctagagcctg ggtttggccagagaattggg tcccggtcag aagtgagtgg ggatggctgg 300 cgagcaaggt gtctgtagggcagcacagga tgtctggtga gcagacagca agcttctgtc 360 ctgccccgag tgctgaggagcgaggtgact gcctacatgg tgatgsaaag atttgggcac 420 gcttccggct ttcaggccaaacaacctcgc ttgctccatg gcaccactga tcccagcagt 480 ggcccgaggg agctccttcctgctgcttca tgctctgaca ctttgggggg ctcctttccc 540 caccacgtgg gtctcctgtcagcctcgaag tgtcctgcgc ccctcncctg tacgcccagg 600 tgtgcctccc ctggccgcacytcctctgtg ctcctgcgtc tctctgttct tctttagagt 660 ggttctgcac gtcagcagcatctgtggtgt ggccctggga cccttcagaa caggggctcc 720 tgcccagctt ctgggtcccccacctgtggc ccagggaagg ctctttgttc ctcagcccca 780 agctgtatct ggtgagaacagatgcgtagt cccggagctc aagttctggg aagggcagtg 840 cccttttctg tggggccctgggcttgttct gcattgtttc aagaggagct gccactcaaa 900 taggcagccc tgcaatcggagggctgcgtg ctccccctga tcagccccca gctgcttcct 960 cgtgcc 966 38 416 DNAHomo sapiens SITE (395) n equals a,t,g, or c 38 gaattcggca cgaggtaataggagccctcg tacctcttgt gttccttaca aacattctca 60 tcagtagctc tacgcgttgactgggtggtt tgaratggct ggtatacaca gggctttctt 120 ggtgttctgt ctctggggcttarctttgtg tgtggttgga gggccctggt gagattggaa 180 gtaccagaga gtgctgtgtcaggggcagag gggcctgtcg ctggagctgg agggtgcctg 240 cctttgtgtc tgactcartctcctgtctgc cttgccccct cagggtctcg ccagcccagc 300 ctctgtggga atctaaaaggartggatgtg gacgtktgac caagcacatc tcagctttta 360 atacctgggc tatttatagacctttggggg gaatngcttg tggaacaaca agggtt 416 39 1114 DNA Homo sapiens 39tgtgtatttg gggggactga agggtacgtg gggcgaaaca aaaccggcca tggcagcagc 60ggaggaggag gacgggggcc ccgaagccaa aatcgcgagc ggggcggggc gggcgcgacc 120ttcgaatgta atatatgttt ggagactgct cgggaagctg tggtcagtgt gtgtggccac 180ctgtactgtt ggccatgtct tcatcagtgg ctggagacac ggccagaacg gcaagagtgt 240ccagtatgta aagctgggat cagcagagag aaggttgtcc cgctttatgg gcgagggagc 300cagaagcccc aggatcccag attaaaaact ccaccccgcc cccagggcca gagaccagct 360ccggagagca gagggggatt ccagccattt ggtgataccg ggggcttcca cttctcattt 420ggtgttggtg cttttccctt tggctttttc accaccgtct tcaatgccca tgagcctttc 480cgccggggta caggtgtgga tctgggacag ggtcacccag cctccagctg gcaggattcc 540ctcttcctgt ttctcgccat cttcttcttt ttttggctgc tcagtatttg agctatgtct 600gcttcctgcc cacctccagc cagagaagaa tcagtattga gggtccctgc tgacccttcc 660gtactcctgg acccccttga cccctctatt tctgttggct aaggccagcc ctggacattg 720tccaggaagg cctggggagg aggagtgaag tctgtgcata gatgggagag ccttctgctc 780agaggctcac tcagtaacgt tgtttaattc tctgccctgg ggaaggagga tggattgaga 840gaatgtcttt ctcctctcct aagtctttgc tttccctgat ttcttgattt gatcttcaaa 900ggtgggcaaa gttccctctg actcttcccc cactccccat cttactgatt taatttaatt 960tttcactccc cagagtctaa tatggattct gactcttaag tgcttccgcc ccctcactac 1020ctcctttaat acaaattcaa taaaaaaggt gaaatataaa aaaaaaaaaa aaaaaacycg 1080gggggggccc cggtccccat tccctttggg gggt 1114 40 602 DNA Homo sapiens SITE(597) n equals a,t,g, or c 40 gggtcgaccc acgcgtccgt cccaggccacaagacatttc ctgctcggaa ccttgtttac 60 taattgtctc tgtggcacat tttgtttcccgtgccttggg tgtcaagttg cagctgatat 120 gaatgaatgc tgtctgtgtg gaacaagcgtcgcaatgagg actctctaca ggacccgata 180 tggcatccct ggatctattt gtgatgactatatggcaact ctttgctgtc ctcattgtac 240 tctttgccaa atcaagagag atatcaacagaaggagagcc atgcgtactt tctaaaaact 300 gatggtgaaa agctcttacc gaagcaacaaaattcagcag acacctcttc agcttgagtt 360 cttcaccatc ttttgcaact gaaatatgatggatatgctt aagtacaact gatggcatga 420 aaaaaatcaa atttttgatt tattataaatgaatgttgtc cctgaactta gctaaatggt 480 gcaacttagt ttctccttgc tttcatattatcgaatttcc tggcttataa actttttaaa 540 ttacatttga aatataaacc aaatgaaatattttactgaa aaaaaaaaaa aaaaaanccc 600 ca 602 41 970 DNA Homo sapiens SITE(37) n equals a,t,g, or c 41 ggcagagctt aggagaacag ctccctttgg atccctntcaaaggtgatac cattggctcc 60 cagcttagag taagaagctc tgagaagttg aatgaagggtgagatagaga tgctgaaccc 120 attcttscag cttcttctag tgttgttatt tccagaatggccaacacccc tacattgata 180 cataaacaca ttccaaggcc ttgtgtaata caaagttcaccgtcctcctg gaataggagc 240 cctgggttct agttctcact ctgccactgg gggaaaatccaattaaagtc tggtttagtc 300 agcttgggtc accatagact gggtggctta aacagcagacatttatttct ggtagtttct 360 ggaggctaca aatctaagag caaggtgcca gcatggtcacattctggtga gggscctctt 420 cctggcttgt agacggctgc yttctcaccg tgtgctcacatagcctttcg tgtgtgtgtg 480 tgtgtgtgtg tgcgtkcgtg caagcttcck gatgtctcttcttagaagga caccaacccc 540 atcatgagag ccctactctc atgacttagc ctaaccctaattaccctcca aaggccccat 600 ctccaaatgc catcacattg gagggtagag cttcaacatagggattttgg gggacacaaa 660 cattcagtcc ataacaaagg ctgtagtcct tartttccttgtctgtgaaa tgagagtgtt 720 gagattcttt ctagccttta tcatttataa ttctgtgagatgtagatttg cattattttc 780 gagttcgagt tatatgaaat gtttccctct acattttcttgggcaactga gaactgaata 840 gggctaggtt taaatagagt taggcagtta ggcttattcttttatttaat aagcattttt 900 ggagcatcta cggtgttcca ggaactgaac tgttgtaaacattggagctg taacagagaa 960 caaaagagac 970 42 1002 DNA Homo sapiens 42gaattcggca cgagccgagg tcggcagcac agagctctgg agatgaagac cctgttcctg 60ggtgtcacgc tcggmctggc cgctgccctg tccttmaccc tggrggagga ggatatcaca 120gggacctggt acgtgaaggc catggtggtc gataagactt tccggagaca ggaggcccag 180aaggtgtccc cagtgaaggt gacagccctg ggcggtggga agttggaagc cacgttcacc 240ttcatgaggg aggatcggtg catccagaag aaaatcctgr tgcggaagac ggaggagcct 300ggcaaataca gcgcctgtga gcccctcccc caytcccacc cccaccytcc cccaccgcca 360accccagtgc accagcctcc acaggtagag agtgcccagg ctgccctttt gccagggccc 420cagctctgcc cacctccaag gaggggctgg cctctccttc ctggggggct ggtggccctg 480acatcagaca ccgggtgtga caggcttgtc cgcagtcgag atggaccaga tcacgcctgc 540cctctgggag gccctagcca ttgacacatt gaggaagctg aggattggga caaggaggcc 600aaggattaga tgggggcagg aagctcatgt acctgcagga gctgcccagg agggaccayt 660acatctttta ctgcaaagac cagcaccatg ggggcstgct ccacatggga aagcttgtgg 720gtaggaattc tgataccaac cgggaggccc tggaagaatt taagaaattg gtgcagcgca 780agggactctc ggaggaggac attttcacgc ccctgcagac gggaagctgc gttcccgaac 840actaggcagc ccccgggtct gcacctccag agcccaccct accaccagac acagagcccg 900gaccacctgg acctaccctc cagccatgac ccttccctgc tcccacccac ctgactccaa 960ataaagtcct tctcccccaa aaaaaaaaaa aaaaaaactc ga 1002 43 2581 DNA Homosapiens SITE (1591) n equals a,t,g, or c 43 tgcaaaacca ctggacactggacaagtacg ggatcctggs cgacgcacgc ctcttctttg 60 ggccccagca ccggsccgtcatccttcggt tgtccaaccg ccgcgcactg cgcctccgtg 120 ccagcttctc ccagcccctcttccaggctg tggstgccat ctgccgcctc ctcagcatcc 180 ggcaccccga ggagctgtccctgctccggg ctcctgagaa gaaggagaag aagaagaaag 240 agaaggagcc agaggaagagctctatgact tgagcaaggt tgtcttggct gggggcgtgg 300 cacctgcact gttccgggggatgccagctc acttctcgga cagcgcccag actgaggcct 360 gctaccacat gctgagccggccccagccgc cacccgaccc cctcctgctc cagcgtctgc 420 cacggcccag ctccctgtcagacaagaccc agctccacag caggtggctg gactcgtcgc 480 ggtgtctcat gcagcagggcatcaaggccg gggacgcact ctggctgcgc ttcaagtact 540 acagcttctt cgatttggatcccaagacag accccgtgcg gctgacacag ctgtatgagc 600 aggcccggtg ggacctgctgctggaggaga ttgactgcac cgaggaggag atgatggtgt 660 ttgccgccct gcagtaccacatcaacaagc tgtcccagag cggggaggtg ggggagccgg 720 ctggcacaga cccagggctggacgacctgg atgtggccct gagcaacctg gaggtgaagc 780 tggaggggtc ggcgcccacagatgtgctgg acagcctcac caccatccca gagctcaagg 840 accatctccg aatctttcggccccggaagc tgaccctgaa gggctaccgc caacactggg 900 tggtgttcaa ggagaccacactgtcctact acaagagcca ggacgaggcc cctggggacc 960 ccattcagca gctcaacctcaagggctgtg aggtggttcc cgatgttaac gtctccggcc 1020 agaagttctg cattaaactcctagtgccct cccctgaggc atgagtgaga tctacctgcg 1080 gtgccaggat gagcagcagtatgcccgctg gatggctggc tgccgcctgg cctccaaagg 1140 ccgcaccatg gccgacagcagctacaccag cgaggtgcag gccatcctgg cyttcctcag 1200 cctgcagcgc acgggcagtgggggcccggg caaccacccc cacggccctg atgcctctgc 1260 cgagggcctc aacccctacggcctcgttgc cccccgtttc cagcgaaagt tcaaggccaa 1320 gcagctcacc ccacggatcctggaagccca ccagaatgtg gcccagttgt cgctggcaga 1380 ggcccagctg cgcttcatccaggcctggca gtccctgccc gacttcggca tctcctatgt 1440 catggtcagg ttcaagggcagcaggaaaga cgagatcctg ggcatcgcca acaaccgact 1500 gatccgcatc gacttggccgtgggcgacgt ggtcaagacc tggcgtttca gcaacatgcg 1560 ccagtggaat gtcaactgggacatccggca ngtggccatc gagtttgatg aacacatcaa 1620 tgtggccttc agctgcgtgtctgccagctg ccgaattgta cacgagtata tcgggggcta 1680 cattttcctg tcgacgcgggagngggcccg tggggaggag ctggatgaag acctcttcct 1740 gcagctcacc gggggccatgaggccttctg agggctgtct gattgcccct gccctgctca 1800 ccaccctgtc acagccactcccaagcccac acccacaggg gctcactgcc ccacacccgc 1860 tccaggcagg cacccagctgggcatttcac ctgctgtcac tgactttgtg caggccaagg 1920 acctggcagg gccagacgctgtaccatcac ccaggccagg gatgggggtg ggggtccctg 1980 agctcatgtg gtgccccctttccttgtctg agtggctgag gctgataccc ctgacctatc 2040 tgcagtcccc cagcacacaaggaagaccag atgtagctac aggatgatga aacatggttt 2100 caaacgagtt ctttcttgttactttttaaa atttcttttt tataaattaa tattttattg 2160 ttggatcctc ctcctttctctggagctgtg cttggggcta ctctgacact ctgtctcttc 2220 atcaccagcc aaggaaaggggctttcctga taaagacaag agttggttag agaaagggac 2280 acctaagtca gtctagggttggaagctagg agagaggtga gggcagaagg gcacagcttt 2340 caggaacaag gaataggggctggggtkgtk gttctcacgg gtaggcggta cctgcagggc 2400 ctccttgaag tacttgggaaggaggaagcc atcagtattc cctggagtca gaatcacccc 2460 attggcagag cggaagaagggtattccatc tgctgacaga gccagagatg tgactcatgc 2520 cctccccgaa ggcaaagtcagctcctgctt tgtccagact cacctgccag agccaggggt 2580 c 2581 44 796 DNA Homosapiens 44 accttcttcc atgtttagtc ccttgggctc tgctaccctc ctgctggaggtgagagcatc 60 ctgtgtgcaa ccagagatgc cctctggctt tcagacctgc ctgcttttcaccctcagccc 120 tttctcactc agcaaaattg tgggggtccc tagtcagcag ctccctgggcagctctctga 180 gcaaggtggt ctctgtggtc atgaaggaga gccggctagg acagtgccggaaactcagct 240 gcctctcccc ttcaactcag ctggcccccc gcacctgaag tgcacaggagccgggaagag 300 agtctggagc ccaccccgga gggcagcaca ggaggtgtct ytgcagctggtgtcctgcma 360 cccytgcagg cagmacacgt cccgggcatt ytcyttagcc acagacagaacagccagtgc 420 cagagtctgc tgtcgyttcc cctttaagca cactcattca ccacacccgaggaggccaga 480 ggtgcaggga gcatgggctg tcgttcccct ttaagcacac tcattcaccacacccgagga 540 ggccagaagt gcagggagca tgggctgggt gcacctccgc aggagagaaggctgagccac 600 cgccgtcccg ggagcccggc tcccaggcct ctcgttttcc cctacctccctaagactttt 660 ctgtcactct ctggccattg aaaggcttct gttccttaaa gtgctgttacactctccttt 720 cccaggatgc agcaagccaa aacagtacca ctgcacgtca gcctgggtgacagagtgaga 780 ccctatctta aaaaaa 796 45 2017 DNA Homo sapiens 45aattcggcac gagcggatcc gttgcggctg cagctctgca gtcgggccgt tccttcgccg 60ccgccagggg tagcggtgta gctgcgcacg tcgcgcgcgc taccgcaccc aggttcggcc 120cgtagcgtct ggcagcccgg cgccatcttc atcgagcgcc atggccgcag cctgcgggcc 180gggagcggcg ggtactgctt gctcctcggc ttgcatttgt ttctgctgac cgcgggccct 240gcctgggctg gaacgaccct gacagaatgt tgctgcggga tgtaaaagct cttaccctcc 300actatgaccg ctataccacc tcccgcagct ggatcccatc ccacagttga aatgtgttgg 360aggcacagct ggttgtgatt cttatacccc aaaagtcata cagtgtcaga acaaaggctg 420ggatgggtat gatgtacagt gggaatgtaa gacggactta gatattgcat acaaatttgg 480aaaaactgtg gtgagctgtg aaggctatga gtcctctgaa gaccagtatg tactaagagg 540ttcttgtggc ttggagtata atttagatta tacagaactt ggcctgcaga aactgaagga 600gtctggaaag cagcacggct ttgcctcttt ctctgattat tattataagt ggtcctcggc 660ggattcctgt aacatgagtg gattgattac catcgtggta ctccttggga tcgcctttgt 720agtctataag ctgttcctga gtgacgggca gtattctcct ccaccgtact ctgagtatcc 780tccattttcc caccgttacc agagattcac caactcagca ggacctcctc ccccaggctt 840taagtctgag ttcacaggac cacagaatac tggccatggt gcaacttctg gttttggcag 900tgcttttaca ggacaacaag gatatgaaaa ttcaggacca gggttctgga caggcttggg 960aactggtgga atactaggat atttgtttgg cagcaataga gcggcaacac ccttctcaga 1020ctcgtggtac tacccgtcct atcctccctc ctaccctggc acgtggaata gggcttactc 1080accccttcat ggaggctcgg gcagctattc ggtatgttca aactcagaca cgaaaaccag 1140aactgcatca ggatatggtg gtaccaggag acgataaagt agaaagttgg agtcaaacac 1200tggatgcaga aattttggat ttttcatcac tttctcttta gaaaaaaagt actacctgtt 1260aacaattggg aaaaggggat attcaaaagt tctgtggtgt tatgtccagt gtagcttttt 1320gtattctatt atttgaggct aaaagttgat gtgtgacaaa atacttatgt gttgtatgtc 1380agtgtaacat gcagatgtat attgcagttt ttgaaagtga tcattactgt ggaatgctaa 1440aaatacatta atttctaaaa cctgtgatgc cctaagaagc attaagaatg aaggtgttgt 1500actaatagaa actaagtaca gaaaatttca gttttaggtg gttgtagctg atgagttatt 1560acctcataga gactataata ttctatttgg tattatatta tttgatgttt gctgttcttc 1620aaacatttaa atcaagcttt ggactaatta tgctaatttg tgagttctga tcacttttga 1680gctctgaagc tttgaatcat tcagtggtgg agatggcctt ctggtaactg aatattacct 1740tctgtaggaa aaggtggaaa ataagcatct agaaggttgt tgtgaatgac tctgtgctgg 1800caaaaatgct tgaaacctct atatttcttt cgttcataag aggtaaaggt caaatttttc 1860aacaaaagtc ttttaataac aaaagcatgc agttctctgt gaaatctcaa atattgttgt 1920aatagtctgt ttcaatctta aaaagaatca ataaaaacaa acaagggaaa aaaaaaaaaa 1980aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 2017 46 981 DNA Homo sapiens 46tcggcagcac agagctctgg agatgaagac cctgttcctg ggtgtcacgc tcggcctggc 60gctgccctgt ccttcaccct ggrggaggag gatatcacag ggacctggta cgtgaaggcc 120atggtggtcg ataagacttt ccggagacag gaggcccaga aggtgtcccc agtgaaggtg 180acagccctgg gcggtgggaa gttggaagcc acgttcacct tcatgaggga ggatcggtgc 240atccagaaga aaatcctgrt gcggaagacg gaggagcctg gcaaatacag cgcctgtgag 300cccctccccc aytcccaccc ccaccytccc ccaccgccaa ccccagtgca ccagcctcca 360caggtagaga gtgcccaggc tgcccttttg ccagggcccc agctctgccc acctccaagg 420aggggctggc ctctccttcc tggggggctg gtggccctga catcagacac cgggtgtgac 480aggcttgtcc gcagtcgaga tggaccagat cacgcctgcc ctctgggagg ccctagccat 540tgacacattg aggaagctga ggattgggac aaggaggcca aggattagat gggggcagga 600agctcatgta cctgcaggag ctgcccagga gggaccayta catcttttac tgcaaagacc 660agcaccatgg gggcstgctc cacatgggaa agcttgtggg taggaattct gataccaacc 720gggaggccct ggaagaattt aagaaattgg tgcagcgcaa gggactctcg gaggaggaca 780ttttcacgcc cctgcagacg ggaagctgcr ttcccgaaca ctaggcagcc cccgggtctg 840cacctccaga gcccacccta ccaccagaca cagagcccgg accacctgga cctaccctcc 900agccatgacc cttccctgct cccacccacc tgactccaaa taaagtcctt ctcccccaaa 960aaaaaaaaaa aaaaaactcg a 981 47 146 PRT Homo sapiens SITE (146) Xaaequals stop translation 47 Met His Tyr Gln Met Ser Val Thr Leu Lys TyrGlu Ile Lys Lys Leu 1 5 10 15 Ile Tyr Val His Leu Val Ile Trp Leu LeuLeu Val Ala Lys Met Ser 20 25 30 Val Gly His Leu Arg Leu Leu Ser His AspGln Val Ala Met Pro Tyr 35 40 45 Gln Trp Glu Tyr Pro Tyr Leu Leu Ser IleLeu Pro Ser Leu Leu Gly 50 55 60 Leu Leu Ser Phe Pro Arg Asn Asn Ile SerTyr Leu Val Leu Ser Met 65 70 75 80 Ile Ser Met Gly Leu Phe Ser Ile AlaPro Leu Ile Tyr Gly Ser Met 85 90 95 Glu Met Phe Pro Ala Ala Gln Pro SerThr Ala Met Ala Arg Pro Thr 100 105 110 Val Ser Ser Leu Val Phe Leu ProPhe Pro Ser Cys Thr Trp Cys Trp 115 120 125 Cys Trp Gln Cys Lys Cys MetPro Gly Ser Cys Thr Thr Ala Arg Ser 130 135 140 Ser Xaa 145 48 312 PRTHomo sapiens SITE (312) Xaa equals stop translation 48 Met Asn Ser ValVal Ser Leu Leu Leu Ile Leu Glu Pro Asp Lys Gln 1 5 10 15 Glu Ala LeuIle Glu Ser Leu Cys Glu Lys Leu Val Lys Phe Arg Glu 20 25 30 Gly Glu ArgPro Ser Leu Arg Leu Gln Leu Leu Ser Asn Leu Phe His 35 40 45 Gly Met AspLys Asn Thr Pro Val Arg Tyr Thr Val Tyr Cys Ser Leu 50 55 60 Ile Lys ValAla Ala Ser Cys Gly Ala Ile Gln Tyr Ile Pro Thr Glu 65 70 75 80 Leu AspGln Val Arg Lys Trp Ile Ser Asp Trp Asn Leu Thr Thr Glu 85 90 95 Lys LysHis Thr Leu Leu Arg Leu Leu Tyr Glu Ala Leu Val Asp Cys 100 105 110 LysLys Ser Asp Ala Ala Ser Lys Val Met Val Glu Leu Leu Gly Ser 115 120 125Tyr Thr Glu Asp Asn Ala Ser Gln Ala Arg Val Asp Ala His Arg Cys 130 135140 Ile Val Arg Ala Leu Lys Asp Pro Asn Ala Phe Leu Phe Asp His Leu 145150 155 160 Leu Thr Leu Lys Pro Val Lys Phe Leu Glu Gly Glu Leu Ile HisAsp 165 170 175 Leu Leu Thr Ile Phe Val Ser Ala Lys Leu Ala Ser Tyr ValLys Phe 180 185 190 Tyr Gln Asn Asn Lys Asp Phe Ile Asp Ser Leu Gly LeuLeu His Glu 195 200 205 Gln Asn Met Ala Lys Met Arg Leu Leu Thr Phe MetGly Met Ala Val 210 215 220 Glu Asn Lys Glu Ile Ser Phe Asp Thr Met GlnGln Glu Leu Gln Ile 225 230 235 240 Gly Ala Asp Asp Val Glu Ala Phe ValIle Asp Ala Val Arg Thr Lys 245 250 255 Met Val Tyr Cys Lys Ile Asp GlnThr Gln Arg Lys Val Val Val Ser 260 265 270 His Ser Thr His Arg Thr PheGly Lys Gln Gln Trp Gln Gln Leu Tyr 275 280 285 Asp Thr Leu Asn Ala TrpLys Gln Asn Leu Asn Lys Val Lys Asn Ser 290 295 300 Leu Leu Ser Leu SerAsp Thr Xaa 305 310 49 64 PRT Homo sapiens 49 Met Met Ser Phe Phe CysPhe Val Met Gly Val Thr Val Ala Ala Thr 1 5 10 15 Phe Thr Ala Ile ValPro Arg Trp Arg Leu Ser Gln Lys Glu Ile Gly 20 25 30 Ser Val Leu Ser ValTrp Leu Ser Arg Trp Arg Glu Asn Ser Leu Arg 35 40 45 Ser Leu Val Ser GlnSer Val Ala Arg Ser Gly Lys Val Val Ile Arg 50 55 60 50 467 PRT Homosapiens 50 Met Leu Ser Arg Pro Gln Pro Pro Pro Asp Pro Leu Leu Leu GlnArg 1 5 10 15 Leu Pro Arg Pro Ser Ser Leu Ser Asp Lys Thr Gln Leu HisSer Arg 20 25 30 Trp Leu Asp Ser Ser Arg Cys Leu Met Gln Gln Gly Ile LysAla Gly 35 40 45 Asp Ala Leu Trp Leu Arg Phe Lys Tyr Tyr Ser Phe Phe AspLeu Asp 50 55 60 Pro Lys Thr Asp Pro Val Arg Leu Thr Gln Leu Tyr Glu GlnAla Arg 65 70 75 80 Trp Asp Leu Leu Leu Glu Glu Ile Asp Cys Thr Glu GluGlu Met Met 85 90 95 Val Phe Ala Ala Leu Gln Tyr His Ile Asn Lys Leu SerGln Ser Gly 100 105 110 Glu Val Gly Glu Pro Ala Gly Thr Asp Pro Gly LeuAsp Asp Leu Asp 115 120 125 Val Ala Leu Ser Asn Leu Glu Val Lys Leu GluGly Ser Ala Pro Thr 130 135 140 Asp Val Leu Asp Ser Leu Thr Thr Ile ProGlu Leu Lys Asp His Leu 145 150 155 160 Arg Ile Phe Arg Pro Arg Lys LeuThr Leu Lys Gly Tyr Arg Gln His 165 170 175 Trp Val Val Phe Lys Glu ThrThr Leu Ser Tyr Tyr Lys Ser Gln Asp 180 185 190 Glu Ala Pro Gly Asp ProIle Gln Gln Leu Asn Leu Lys Gly Cys Glu 195 200 205 Val Val Pro Asp ValAsn Val Ser Gly Gln Lys Phe Cys Ile Lys Leu 210 215 220 Leu Val Pro SerPro Glu Gly Met Ser Glu Ile Tyr Leu Arg Cys Gln 225 230 235 240 Asp GluGln Gln Tyr Ala Arg Trp Met Ala Gly Cys Arg Leu Ala Ser 245 250 255 LysGly Arg Thr Met Ala Asp Ser Ser Tyr Thr Ser Glu Val Gln Ala 260 265 270Ile Leu Ala Phe Leu Ser Leu Gln Arg Thr Gly Ser Gly Gly Pro Gly 275 280285 Asn His Pro His Gly Pro Asp Ala Ser Ala Glu Gly Leu Asn Pro Tyr 290295 300 Gly Leu Val Ala Pro Arg Phe Gln Arg Lys Phe Lys Ala Lys Gln Leu305 310 315 320 Thr Pro Arg Ile Leu Glu Ala His Gln Asn Val Ala Gln LeuSer Leu 325 330 335 Ala Glu Ala Gln Leu Arg Phe Ile Gln Ala Trp Gln SerLeu Pro Asp 340 345 350 Phe Gly Ile Ser Tyr Val Met Val Arg Phe Lys GlySer Arg Lys Asp 355 360 365 Glu Ile Leu Gly Ile Ala Asn Asn Arg Leu IleArg Ile Asp Leu Ala 370 375 380 Val Gly Asp Val Val Lys Thr Trp Arg PheSer Asn Met Arg Gln Trp 385 390 395 400 Asn Val Asn Trp Asp Ile Arg GlnVal Ala Ile Glu Phe Asp Glu His 405 410 415 Ile Asn Val Ala Phe Ser CysVal Ser Ala Ser Cys Arg Ile Val His 420 425 430 Glu Tyr Ile Gly Gly TyrIle Phe Leu Ser Thr Arg Glu Arg Ala Arg 435 440 445 Gly Glu Glu Leu AspGlu Asp Leu Phe Leu Gln Leu Thr Gly Gly His 450 455 460 Glu Ala Phe 46551 83 PRT Homo sapiens SITE (83) Xaa equals stop translation 51 Met ArgPro Gly Arg Gly Ala Gly Thr Pro Gly Arg Pro Gly Arg Gly 1 5 10 15 ArgGly Leu Ala Ala Thr Cys Ser Leu Ser Ser Pro Ser His Leu Leu 20 25 30 ProThr Leu Leu His Thr Phe Ser Phe Ser Leu Pro Pro Pro Ser Pro 35 40 45 AlaAla Pro Arg Gln Pro Ser Pro Pro Ala Leu Leu Leu Pro Gly Pro 50 55 60 GlnLys Pro Arg Pro Gly Asp Pro Thr Tyr Thr Gly Ala Leu Thr Asp 65 70 75 80Trp Ser Xaa 52 63 PRT Homo sapiens SITE (63) Xaa equals stop translation52 Met Phe Leu Val Phe Phe Leu Ser Phe Phe Ser His Ser Ile Ser Ala 1 510 15 Leu Thr Leu Val Cys Ser Gln Gly Gly Lys Ala Asp Met Asn Leu Leu 2025 30 Ser Trp Asp Phe Arg Pro His Trp Leu Glu Gly Ile Arg Phe Leu Leu 3540 45 Gly Trp Gly Gln Ala Leu Met Ala Gly Leu Phe Pro Trp Leu Xaa 50 5560 53 124 PRT Homo sapiens SITE (114) Xaa equals any of the naturallyoccurring L- amino acids 53 Met Arg Gly Ser Trp His Arg Ser Pro Leu ProAla Val Val Leu Pro 1 5 10 15 Ser Val Leu Gln Thr Ala Leu Ser Pro LeuAla Leu Cys Gln Ala Trp 20 25 30 Arg Arg Ala Val Pro His Gly Val Pro SerGln Arg Leu Arg Asn Gln 35 40 45 Glu Ala Ser Leu Val Pro Lys Gly Val ProArg Ala Trp Tyr Pro Gly 50 55 60 Pro Leu Gln Asn Gly Leu Trp Thr His LeuGlu Lys Gly Glu Leu Leu 65 70 75 80 Gly Leu Lys Pro Thr Pro Gly Gly LeuLeu Leu Leu Arg Ser Phe Trp 85 90 95 Asp Pro His Pro Ser Arg Pro Phe LeuCys Thr Leu Leu Pro Pro Pro 100 105 110 Leu Xaa Ile Phe Pro Pro Leu ArgCys Ser Ala Xaa 115 120 54 180 PRT Homo sapiens SITE (8) Xaa equals anyof the naturally occurring L- amino acids 54 Met Thr Ser Ala Gly Pro ValXaa Leu Phe Leu Leu Val Ser Ile Ser 1 5 10 15 Thr Ser Val Ile Leu MetGln His Leu Leu Xaa Ala Ser Tyr Cys Asp 20 25 30 Leu Leu His Lys Ala AlaAla His Leu Gly Cys Trp Gln Lys Val Asp 35 40 45 Pro Ala Leu Cys Ser AsnVal Leu Gln His Pro Trp Thr Glu Glu Cys 50 55 60 Met Trp Pro Gln Gly ValLeu Val Lys His Ser Lys Asn Val Tyr Lys 65 70 75 80 Ala Val Gly Xaa XaaXaa Val Ala Ile Pro Ser Asp Val Ser His Phe 85 90 95 Arg Phe Xaa Phe PhePhe Ser Lys Pro Leu Arg Ile Leu Asn Ile Leu 100 105 110 Leu Leu Leu GluGly Ala Val Ile Val Tyr Gln Leu Tyr Ser Leu Met 115 120 125 Ser Ser GluLys Trp His Gln Thr Ile Ser Leu Ala Leu Ile Leu Phe 130 135 140 Ser AsnTyr Tyr Ala Phe Phe Lys Leu Leu Arg Asp Arg Leu Val Leu 145 150 155 160Gly Lys Ala Tyr Ser Tyr Ser Ala Ser Pro Gln Arg Asp Leu Asp His 165 170175 Arg Phe Ser Xaa 180 55 287 PRT Homo sapiens SITE (221) Xaa equalsany of the naturally occurring L- amino acids 55 Met Pro Leu Phe Lys LeuTyr Met Val Met Ser Ala Cys Phe Leu Ala 1 5 10 15 Ala Gly Ile Phe TrpVal Ser Ile Leu Cys Arg Asn Thr Tyr Ser Val 20 25 30 Phe Lys Ile His TrpLeu Met Ala Ala Leu Ala Phe Thr Lys Ser Ile 35 40 45 Ser Leu Leu Phe HisSer Ile Asn Tyr Tyr Phe Ile Asn Ser Gln Gly 50 55 60 Pro Pro His Arg ArgPro Cys Arg His Val Leu His Arg Thr Pro Ala 65 70 75 80 Glu Gly Arg ProPro Leu His His His Arg Pro Asp Trp Leu Arg Leu 85 90 95 Gly Phe Ile LysTyr Val Leu Ser Asp Lys Glu Lys Lys Val Phe Gly 100 105 110 Ile Val IlePro Met Gln Val Leu Ala Asn Val Ala Tyr Ile Ile Ile 115 120 125 Glu SerArg Glu Glu Gly Ala Thr Asn Tyr Val Leu Trp Lys Glu Ile 130 135 140 LeuPhe Leu Val Asp Leu Ile Cys Cys Gly Ala Ile Leu Phe Pro Val 145 150 155160 Val Trp Ser Ile Arg His Leu Gln Asp Ala Ser Gly Thr Asp Gly Lys 165170 175 Val Ala Val Asn Leu Ala Lys Leu Lys Leu Phe Arg His Tyr Tyr Val180 185 190 Met Val Ile Cys Tyr Val Tyr Phe Thr Arg Ile Ile Ala Ile LeuLeu 195 200 205 Gln Val Ala Val Pro Phe Gln Trp Gln Trp Leu Tyr Xaa LeuLeu Val 210 215 220 Glu Gly Ser Thr Leu Ala Phe Phe Val Leu Thr Gly TyrLys Phe Gln 225 230 235 240 Pro Thr Gly Asn Asn Pro Tyr Leu Gln Leu ProGln Glu Asp Glu Glu 245 250 255 Asp Val Gln Met Glu Gln Val Met Thr AspSer Gly Phe Arg Glu Gly 260 265 270 Leu Ser Lys Val Asn Lys Thr Ala SerGly Arg Glu Leu Leu Xaa 275 280 285 56 34 PRT Homo sapiens SITE (34) Xaaequals stop translation 56 Met Pro Met Val Phe Leu Leu Leu Phe Asn LeuMet Ser Trp Leu Ile 1 5 10 15 Arg Asn Ala Arg Val Ile Leu Arg Ser LeuAsn Leu Lys Arg Asp Gln 20 25 30 Val Xaa 57 24 PRT Homo sapiens SITE(24) Xaa equals stop translation 57 Met Lys Ile Val Val Leu Leu Pro LeuPhe Leu Leu Ala Thr Phe Pro 1 5 10 15 Arg Lys Leu Gln Thr Cys Leu Xaa 2058 47 PRT Homo sapiens SITE (47) Xaa equals stop translation 58 Met SerGly Gly Glu Gly Ala Ala Leu Pro Ile Leu Leu Leu Leu Leu 1 5 10 15 AlaLeu Arg Gly Thr Phe His Gly Ala Arg Pro Gly Gly Gly Ala Ser 20 25 30 GlyIle Trp Cys Leu Leu Leu Pro Glu Gln Glu Pro Pro Val Xaa 35 40 45 59 114PRT Homo sapiens SITE (114) Xaa equals stop translation 59 Met Ala ArgGly Ser Leu Arg Arg Leu Leu Arg Leu Leu Val Leu Gly 1 5 10 15 Leu TrpLeu Ala Leu Leu Arg Ser Val Ala Gly Glu Gln Ala Pro Gly 20 25 30 Thr AlaPro Cys Ser Arg Gly Ser Ser Trp Ser Ala Asp Leu Asp Lys 35 40 45 Cys MetAsp Cys Ala Ser Cys Arg Ala Arg Pro His Ser Asp Phe Cys 50 55 60 Leu GlyCys Ala Ala Ala Pro Pro Ala Pro Phe Arg Leu Leu Trp Pro 65 70 75 80 IleLeu Gly Gly Ala Leu Ser Leu Thr Phe Val Leu Gly Leu Leu Ser 85 90 95 GlyPhe Leu Val Trp Arg Arg Cys Arg Arg Glu Arg Ser Ser Pro Pro 100 105 110Pro Xaa 60 32 PRT Homo sapiens SITE (26) Xaa equals any of the naturallyoccurring L- amino acids 60 Met Val Cys Ile Leu Val Leu Thr Leu Val SerTyr Ser Ser Leu Val 1 5 10 15 Asn Ser Pro Leu Pro Phe Val His Leu XaaVal Gly Ile Ser Ala Xaa 20 25 30 61 81 PRT Homo sapiens SITE (19) Xaaequals any of the naturally occurring L- amino acids 61 Met Thr Gly GlyPhe Leu Ser Cys Ile Leu Gly Leu Val Leu Pro Leu 1 5 10 15 Ala Tyr XaaSer Ser Leu Thr Trp Cys Trp Trp Arg Trp Gly Leu Pro 20 25 30 Xaa Pro AlaGly Pro Pro Arg Cys Thr Pro Gly Cys Asn Ala Ser Gly 35 40 45 Ala Gly ArgGly Pro Ser Pro Gly Pro Pro Gly Gly Glu Leu His Thr 50 55 60 Pro Ala SerArg Asp Pro Gly Pro Gly Ala Glu Trp Arg Gly Thr Ser 65 70 75 80 Xaa 62104 PRT Homo sapiens 62 Met Ala Ala Pro Val Asp Leu Glu Leu Lys Lys AlaPhe Thr Glu Leu 1 5 10 15 Gln Ala Lys Val Ile Asp Thr Gln Gln Lys ValLys Leu Ala Asp Ile 20 25 30 Gln Ile Glu Gln Leu Asn Arg Thr Lys Lys HisAla His Leu Thr Asp 35 40 45 Thr Glu Ile Met Thr Leu Val Asp Glu Thr AsnMet Tyr Glu Gly Val 50 55 60 Gly Arg Met Phe Ile Leu Gln Ser Lys Glu AlaIle His Ser Gln Leu 65 70 75 80 Leu Glu Lys Gln Lys Ile Ala Glu Glu LysIle Lys Glu Leu Glu Gln 85 90 95 Lys Lys Ser Tyr Leu Glu Arg Arg 100 63146 PRT Homo sapiens SITE (146) Xaa equals stop translation 63 Met ProSer Gly Phe Gln Thr Cys Leu Leu Phe Thr Leu Ser Pro Phe 1 5 10 15 SerLeu Ser Lys Ile Val Gly Val Pro Ser Gln Gln Leu Pro Gly Gln 20 25 30 LeuSer Glu Gln Gly Gly Leu Cys Gly His Glu Gly Glu Pro Ala Arg 35 40 45 ThrVal Pro Glu Thr Gln Leu Pro Leu Pro Phe Asn Ser Ala Gly Pro 50 55 60 ProHis Leu Lys Cys Thr Gly Ala Gly Lys Arg Val Trp Ser Pro Pro 65 70 75 80Arg Arg Ala Ala Gln Glu Val Ser Leu Gln Leu Val Ser Cys His Pro 85 90 95Cys Arg Gln His Thr Ser Arg Ala Phe Ser Leu Ala Thr Asp Arg Thr 100 105110 Ala Ser Ala Arg Val Cys Cys Arg Ser Pro Leu Ser Thr Leu Ile His 115120 125 His Thr Arg Gly Gly Gln Arg Cys Arg Glu His Gly Leu Ser Leu Pro130 135 140 Leu Xaa 145 64 31 PRT Homo sapiens SITE (31) Xaa equals stoptranslation 64 Met Ala Ile Leu Met Leu Leu Ala Gly Ser Pro Cys Thr LeuSer Phe 1 5 10 15 Ser Thr Asp Thr Gly Ser Ser Ala Pro Gly Pro Lys IlePro Xaa 20 25 30 65 260 PRT Homo sapiens SITE (260) Xaa equals stoptranslation 65 Met Asp Pro Gln Gly Gln Thr Leu Leu Leu Phe Leu Phe ValAsp Phe 1 5 10 15 His Ser Ala Phe Pro Val Gln Gln Met Glu Ile Trp GlyVal Tyr Thr 20 25 30 Leu Leu Thr Thr His Leu Asn Ala Ile Leu Val Glu SerHis Ser Val 35 40 45 Val Gln Gly Ser Ile Gln Phe Thr Val Asp Lys Val LeuGlu Gln His 50 55 60 His Gln Ala Ala Lys Ala Gln Gln Lys Leu Gln Ala SerLeu Ser Val 65 70 75 80 Ala Val Asn Ser Ile Met Ser Ile Leu Thr Gly SerThr Arg Ser Ser 85 90 95 Phe Arg Lys Met Cys Leu Gln Thr Leu Gln Ala AlaAsp Thr Gln Glu 100 105 110 Phe Arg Thr Lys Leu His Lys Val Phe Arg GluIle Thr Gln His Gln 115 120 125 Phe Leu His His Cys Ser Cys Glu Val LysGln Leu Thr Leu Glu Lys 130 135 140 Lys Asp Ser Ala Gln Gly Thr Glu AspAla Pro Asp Asn Ser Ser Leu 145 150 155 160 Glu Leu Leu Ala Asp Thr SerGly Gln Ala Glu Asn Lys Arg Leu Lys 165 170 175 Arg Gly Ser Pro Arg IleGlu Glu Met Arg Ala Leu Arg Ser Ala Arg 180 185 190 Ala Pro Ser Pro SerGlu Ala Ala Pro Arg Arg Pro Glu Ala Thr Ala 195 200 205 Ala Pro Leu ThrPro Arg Gly Arg Glu His Arg Glu Ala His Gly Arg 210 215 220 Ala Leu AlaPro Gly Arg Ala Ser Leu Gly Ser Arg Leu Glu Asp Val 225 230 235 240 LeuTrp Leu Gln Glu Val Ser Asn Leu Ser Glu Trp Leu Ser Pro Ser 245 250 255Pro Gly Pro Xaa 260 66 339 PRT Homo sapiens 66 Met Ala Ala Ala Cys GlyPro Gly Ala Ala Gly Tyr Cys Leu Leu Leu 1 5 10 15 Gly Leu His Leu PheLeu Leu Thr Ala Gly Pro Ala Leu Gly Trp Asn 20 25 30 Asp Pro Asp Arg MetLeu Leu Arg Asp Val Lys Ala Leu Thr Leu His 35 40 45 Tyr Asp Arg Tyr ThrThr Ser Arg Arg Leu Asp Pro Ile Pro Gln Leu 50 55 60 Lys Cys Val Gly GlyThr Ala Gly Cys Asp Ser Tyr Thr Pro Lys Val 65 70 75 80 Ile Gln Cys GlnAsn Lys Gly Trp Asp Gly Tyr Asp Val Gln Trp Glu 85 90 95 Cys Lys Thr AspLeu Asp Ile Ala Tyr Lys Phe Gly Lys Thr Val Val 100 105 110 Ser Cys GluGly Tyr Glu Ser Ser Glu Asp Gln Tyr Val Leu Arg Gly 115 120 125 Ser CysGly Leu Glu Tyr Asn Leu Asp Tyr Thr Glu Leu Gly Leu Gln 130 135 140 LysLeu Lys Glu Ser Gly Lys Gln His Gly Phe Ala Ser Phe Ser Asp 145 150 155160 Tyr Tyr Tyr Lys Trp Ser Ser Ala Asp Ser Cys Asn Met Ser Gly Leu 165170 175 Ile Thr Ile Val Val Leu Leu Gly Ile Ala Phe Val Val Tyr Lys Leu180 185 190 Phe Leu Ser Asp Gly Gln Tyr Ser Pro Pro Pro Tyr Ser Glu TyrPro 195 200 205 Pro Phe Ser His Arg Tyr Gln Arg Phe Thr Asn Ser Ala GlyPro Pro 210 215 220 Pro Pro Gly Phe Lys Ser Glu Phe Thr Gly Pro Gln AsnThr Gly His 225 230 235 240 Gly Ala Thr Ser Gly Phe Gly Ser Ala Phe ThrGly Gln Gln Gly Tyr 245 250 255 Glu Asn Ser Gly Pro Gly Phe Trp Thr GlyLeu Gly Thr Gly Gly Ile 260 265 270 Leu Gly Tyr Leu Phe Gly Ser Asn ArgAla Ala Thr Pro Phe Ser Asp 275 280 285 Ser Trp Tyr Tyr Pro Ser Tyr ProPro Ser Tyr Pro Gly Thr Trp Asn 290 295 300 Arg Ala Tyr Ser Pro Leu HisGly Gly Ser Gly Ser Tyr Ser Val Cys 305 310 315 320 Ser Asn Ser Asp ThrLys Thr Arg Thr Ala Ser Gly Tyr Gly Gly Thr 325 330 335 Arg Arg Arg 6727 PRT Homo sapiens SITE (27) Xaa equals stop translation 67 Met His AlaLeu Ile Leu Gln Phe Ile Phe Ser Leu Cys Met Tyr Ile 1 5 10 15 Ser LeuPhe Ser Ala Ala Arg Phe Leu Phe Xaa 20 25 68 76 PRT Homo sapiens SITE(64) Xaa equals any of the naturally occurring L- amino acids 68 Met SerGln Ser Val Ser Ser Ser Phe Leu Ile Leu Thr Leu Leu Leu 1 5 10 15 SerVal Gly Phe Gln Cys Leu Thr Leu Tyr Thr Thr Val Thr Thr Thr 20 25 30 CysLeu Trp Gly Pro Pro Arg Ala Ala Gly Arg Leu Phe Val Gln Ser 35 40 45 LeuPro Ser Cys Glu Cys Cys Cys Arg Ala Arg Arg Gly Ala Val Xaa 50 55 60 XaaSer Pro Pro Trp Arg Pro Trp Pro Glu Gln Val 65 70 75 69 216 PRT Homosapiens SITE (216) Xaa equals stop translation 69 Met Tyr Leu Ser IleIle Phe Leu Ala Phe Val Ser Ile Asp Arg Cys 1 5 10 15 Leu Gln Leu ThrHis Ser Cys Lys Ile Tyr Arg Ile Gln Glu Pro Gly 20 25 30 Phe Ala Lys MetIle Ser Thr Val Val Trp Leu Met Val Leu Leu Ile 35 40 45 Met Val Pro AsnMet Met Ile Pro Ile Lys Asp Ile Lys Glu Lys Ser 50 55 60 Asn Val Gly CysMet Glu Phe Lys Lys Glu Phe Gly Arg Asn Trp His 65 70 75 80 Leu Leu ThrAsn Phe Ile Cys Val Ala Ile Phe Leu Asn Phe Ser Ala 85 90 95 Ile Ile LeuIle Ser Asn Cys Leu Val Ile Arg Gln Leu Tyr Arg Asn 100 105 110 Lys AspAsn Glu Asn Tyr Pro Asn Val Lys Lys Ala Leu Ile Asn Ile 115 120 125 LeuLeu Val Thr Thr Gly Tyr Ile Ile Cys Phe Val Pro Tyr His Ile 130 135 140Val Arg Ile Pro Tyr Thr Leu Ser Gln Thr Glu Val Ile Thr Asp Cys 145 150155 160 Ser Thr Arg Ile Ser Leu Phe Lys Ala Lys Glu Ala Thr Leu Leu Leu165 170 175 Ala Val Ser Asn Leu Cys Phe Asp Pro Ile Leu Tyr Tyr His LeuSer 180 185 190 Lys Ala Phe Arg Ser Lys Val Thr Glu Thr Phe Ala Ser ProLys Glu 195 200 205 Thr Lys Val Arg Lys Lys Asn Xaa 210 215 70 407 PRTHomo sapiens SITE (407) Xaa equals stop translation 70 Met His Pro AlaVal Phe Leu Ser Leu Pro Asp Leu Arg Cys Ser Leu 1 5 10 15 Leu Leu LeuVal Thr Trp Val Phe Thr Pro Val Thr Thr Glu Ile Thr 20 25 30 Ser Leu AspThr Glu Asn Ile Asp Glu Ile Leu Asn Asn Ala Asp Val 35 40 45 Ala Leu ValAsn Phe Tyr Ala Asp Trp Cys Arg Phe Ser Gln Met Leu 50 55 60 His Pro IlePhe Glu Glu Ala Ser Asp Val Ile Lys Glu Glu Phe Pro 65 70 75 80 Asn GluAsn Gln Val Val Phe Ala Arg Val Asp Cys Asp Gln His Ser 85 90 95 Asp IleAla Gln Arg Tyr Arg Ile Ser Lys Tyr Pro Thr Leu Lys Leu 100 105 110 PheArg Asn Gly Met Met Met Lys Arg Glu Tyr Arg Gly Gln Arg Ser 115 120 125Val Lys Ala Leu Ala Asp Tyr Ile Arg Gln Gln Lys Ser Asp Pro Ile 130 135140 Gln Glu Ile Arg Asp Leu Ala Glu Ile Thr Thr Leu Asp Arg Ser Lys 145150 155 160 Arg Asn Ile Ile Gly Tyr Phe Glu Gln Lys Asp Ser Asp Asn TyrArg 165 170 175 Val Phe Glu Arg Val Ala Asn Ile Leu His Asp Asp Cys AlaPhe Leu 180 185 190 Ser Ala Phe Gly Asp Val Ser Lys Pro Glu Arg Tyr SerGly Asp Asn 195 200 205 Ile Ile Tyr Lys Pro Pro Gly His Ser Ala Pro AspMet Val Tyr Leu 210 215 220 Gly Ala Met Thr Asn Phe Asp Val Thr Tyr AsnTrp Ile Gln Asp Lys 225 230 235 240 Cys Val Pro Leu Val Arg Glu Ile ThrPhe Glu Asn Gly Glu Glu Leu 245 250 255 Thr Glu Glu Gly Leu Pro Phe LeuIle Leu Phe His Met Lys Glu Asp 260 265 270 Thr Glu Ser Leu Glu Ile PheGln Asn Glu Val Ala Arg Gln Leu Ile 275 280 285 Ser Glu Lys Gly Thr IleAsn Phe Leu His Ala Asp Cys Asp Lys Phe 290 295 300 Arg His Pro Leu LeuHis Ile Gln Lys Thr Pro Ala Asp Cys Pro Val 305 310 315 320 Ile Ala IleAsp Ser Phe Arg His Met Tyr Val Phe Gly Asp Phe Lys 325 330 335 Asp ValLeu Ile Pro Gly Lys Leu Lys Gln Phe Val Phe Asp Leu His 340 345 350 SerGly Lys Leu His Arg Glu Phe His His Gly Pro Asp Pro Thr Asp 355 360 365Thr Ala Pro Gly Glu Gln Ala Gln Asp Val Ala Ser Ser Pro Pro Glu 370 375380 Ser Ser Phe Gln Lys Leu Ala Pro Ser Glu Tyr Arg Tyr Thr Leu Leu 385390 395 400 Arg Asp Arg Asp Glu Leu Xaa 405 71 45 PRT Homo sapiens 71Met Ser Met Cys Ile His Ala Lys Lys His Leu Ile Cys Ile Cys Phe 1 5 1015 Arg Lys Gly Gly Asn Glu Ala Thr Cys Leu Lys Ile Leu Leu Tyr Lys 20 2530 Ala Phe Gln Pro Phe Pro Leu Ser Phe Ala Leu Ile Phe 35 40 45 72 34PRT Homo sapiens SITE (34) Xaa equals stop translation 72 Met Pro LeuLys Ala Val Thr Trp Pro Thr Leu Asn Ser Lys Leu Val 1 5 10 15 Ala AlaVal Val Asn Leu Lys Ala Ser Gln Met Pro Ala Ser Ser Arg 20 25 30 Val Xaa73 160 PRT Homo sapiens SITE (55) Xaa equals any of the naturallyoccurring L- amino acids 73 Met Ala Pro Leu Ile Pro Ala Val Ala Arg GlySer Ser Phe Leu Leu 1 5 10 15 Leu His Ala Leu Thr Leu Trp Gly Ala ProPhe Pro Thr Thr Trp Val 20 25 30 Ser Cys Gln Pro Arg Ser Val Leu Arg ProSer Pro Val Arg Pro Gly 35 40 45 Val Pro Pro Leu Ala Ala Xaa Pro Leu CysSer Cys Val Ser Leu Phe 50 55 60 Phe Phe Arg Val Val Leu His Val Ser SerIle Cys Gly Val Ala Leu 65 70 75 80 Gly Pro Phe Arg Thr Gly Ala Pro AlaGln Leu Leu Gly Pro Pro Pro 85 90 95 Val Ala Gln Gly Arg Leu Phe Val ProGln Pro Gln Ala Val Ser Gly 100 105 110 Glu Asn Arg Cys Val Val Pro GluLeu Lys Phe Trp Glu Gly Gln Cys 115 120 125 Pro Phe Leu Trp Gly Pro GlyLeu Val Leu His Cys Phe Lys Arg Ser 130 135 140 Cys His Ser Asn Arg GlnPro Cys Asn Arg Arg Ala Ala Cys Ser Pro 145 150 155 160 74 26 PRT Homosapiens SITE (17) Xaa equals any of the naturally occurring L- aminoacids 74 Met Ala Gly Ile His Arg Ala Phe Leu Val Phe Cys Leu Trp Gly Leu1 5 10 15 Xaa Leu Cys Val Val Gly Gly Pro Trp Xaa 20 25 75 91 PRT Homosapiens 75 Met Ala Ala Ala Glu Glu Glu Asp Gly Gly Pro Glu Ala Lys IleAla 1 5 10 15 Ser Gly Ala Gly Arg Ala Arg Pro Ser Asn Val Ile Tyr ValTrp Arg 20 25 30 Leu Leu Gly Lys Leu Trp Ser Val Cys Val Ala Thr Cys ThrVal Gly 35 40 45 His Val Phe Ile Ser Gly Trp Arg His Gly Gln Asn Gly LysSer Val 50 55 60 Gln Tyr Val Lys Leu Gly Ser Ala Glu Arg Arg Leu Ser ArgPhe Met 65 70 75 80 Gly Glu Gly Ala Arg Ser Pro Arg Ile Pro Asp 85 90 7633 PRT Homo sapiens SITE (33) Xaa equals stop translation 76 Met Thr IleTrp Gln Leu Phe Ala Val Leu Ile Val Leu Phe Ala Lys 1 5 10 15 Ser ArgGlu Ile Ser Thr Glu Gly Glu Pro Cys Val Leu Ser Lys Asn 20 25 30 Xaa 7723 PRT Homo sapiens SITE (6) Xaa equals any of the naturally occurringL- amino acids 77 Met Leu Asn Pro Phe Xaa Gln Leu Leu Leu Val Leu LeuPhe Pro Glu 1 5 10 15 Trp Pro Thr Pro Leu His Xaa 20 78 173 PRT Homosapiens SITE (18) Xaa equals any of the naturally occurring L- aminoacids 78 Met Lys Thr Leu Phe Leu Gly Val Thr Leu Gly Leu Ala Ala Ala Leu1 5 10 15 Ser Xaa Thr Leu Xaa Glu Glu Asp Ile Thr Gly Thr Trp Tyr ValLys 20 25 30 Ala Met Val Val Asp Lys Thr Phe Arg Arg Gln Glu Ala Gln LysVal 35 40 45 Ser Pro Val Lys Val Thr Ala Leu Gly Gly Gly Lys Leu Glu AlaThr 50 55 60 Phe Thr Phe Met Arg Glu Asp Arg Cys Ile Gln Lys Lys Ile LeuXaa 65 70 75 80 Arg Lys Thr Glu Glu Pro Gly Lys Tyr Ser Ala Cys Glu ProLeu Pro 85 90 95 His Ser His Pro His Xaa Pro Pro Pro Pro Thr Pro Val HisGln Pro 100 105 110 Pro Gln Val Glu Ser Ala Gln Ala Ala Leu Leu Pro GlyPro Gln Leu 115 120 125 Cys Pro Pro Pro Arg Arg Gly Trp Pro Leu Leu ProGly Gly Leu Val 130 135 140 Ala Leu Thr Ser Asp Thr Gly Cys Asp Arg LeuVal Arg Ser Arg Asp 145 150 155 160 Gly Pro Asp His Ala Cys Pro Leu GlyGly Pro Ser His 165 170 79 208 PRT Homo sapiens SITE (148) Xaa equalsany of the naturally occurring L- amino acids 79 Met Ala Asp Ser Ser TyrThr Ser Glu Val Gln Ala Ile Leu Ala Phe 1 5 10 15 Leu Ser Leu Gln ArgThr Gly Ser Gly Gly Pro Gly Asn His Pro His 20 25 30 Gly Pro Asp Ala SerAla Glu Gly Leu Asn Pro Tyr Gly Leu Val Ala 35 40 45 Pro Arg Phe Gln ArgLys Phe Lys Ala Lys Gln Leu Thr Pro Arg Ile 50 55 60 Leu Glu Ala His GlnAsn Val Ala Gln Leu Ser Leu Ala Glu Ala Gln 65 70 75 80 Leu Arg Phe IleGln Ala Trp Gln Ser Leu Pro Asp Phe Gly Ile Ser 85 90 95 Tyr Val Met ValArg Phe Lys Gly Ser Arg Lys Asp Glu Ile Leu Gly 100 105 110 Ile Ala AsnAsn Arg Leu Ile Arg Ile Asp Leu Ala Val Gly Asp Val 115 120 125 Val LysThr Trp Arg Phe Ser Asn Met Arg Gln Trp Asn Val Asn Trp 130 135 140 AspIle Arg Xaa Val Ala Ile Glu Phe Asp Glu His Ile Asn Val Ala 145 150 155160 Phe Ser Cys Val Ser Ala Ser Cys Arg Ile Val His Glu Tyr Ile Gly 165170 175 Gly Tyr Ile Phe Leu Ser Thr Arg Glu Xaa Ala Arg Gly Glu Glu Leu180 185 190 Asp Glu Asp Leu Phe Leu Gln Leu Thr Gly Gly His Glu Ala PheXaa 195 200 205 80 146 PRT Homo sapiens SITE (95) Xaa equals any of thenaturally occurring L- amino acids 80 Met Pro Ser Gly Phe Gln Thr CysLeu Leu Phe Thr Leu Ser Pro Phe 1 5 10 15 Ser Leu Ser Lys Ile Val GlyVal Pro Ser Gln Gln Leu Pro Gly Gln 20 25 30 Leu Ser Glu Gln Gly Gly LeuCys Gly His Glu Gly Glu Pro Ala Arg 35 40 45 Thr Val Pro Glu Thr Gln LeuPro Leu Pro Phe Asn Ser Ala Gly Pro 50 55 60 Pro His Leu Lys Cys Thr GlyAla Gly Lys Arg Val Trp Ser Pro Pro 65 70 75 80 Arg Arg Ala Ala Gln GluVal Ser Leu Gln Leu Val Ser Cys Xaa Pro 85 90 95 Cys Arg Gln Xaa Thr SerArg Ala Phe Ser Leu Ala Thr Asp Arg Thr 100 105 110 Ala Ser Ala Arg ValCys Cys Arg Phe Pro Phe Lys His Thr His Ser 115 120 125 Pro His Pro ArgArg Pro Glu Val Gln Gly Ala Trp Ala Val Val Pro 130 135 140 Leu Xaa 14581 23 PRT Homo sapiens SITE (23) Xaa equals stop translation 81 Met AlaAla Ala Cys Gly Pro Gly Ala Ala Gly Thr Ala Cys Ser Ser 1 5 10 15 AlaCys Ile Cys Phe Cys Xaa 20 82 31 PRT Homo sapiens SITE (21) Xaa equalsany of the naturally occurring L- amino acids 82 Met Lys Thr Leu Phe LeuGly Val Thr Leu Gly Leu Ala Leu Pro Cys 1 5 10 15 Pro Ser Pro Trp XaaArg Arg Ile Ser Gln Gly Pro Gly Thr Xaa 20 25 30 83 374 PRT Homo sapiens83 Met Ser Val Pro Ala Phe Ile Asp Ile Ser Glu Glu Asp Gln Ala Ala 1 510 15 Glu Leu Arg Ala Tyr Leu Lys Ser Lys Gly Ala Glu Ile Ser Glu Glu 2025 30 Asn Ser Glu Gly Gly Leu His Val Asp Leu Ala Gln Ile Ile Glu Ala 3540 45 Cys Asp Val Cys Leu Lys Glu Asp Asp Lys Asp Val Glu Ser Val Met 5055 60 Asn Ser Val Val Ser Leu Leu Leu Ile Leu Glu Pro Asp Lys Gln Glu 6570 75 80 Ala Leu Ile Glu Ser Leu Cys Glu Lys Leu Val Lys Phe Arg Glu Gly85 90 95 Glu Arg Pro Ser Leu Arg Leu Gln Leu Leu Ser Asn Leu Phe His Gly100 105 110 Met Asp Lys Asn Thr Pro Val Arg Tyr Thr Val Tyr Cys Ser LeuIle 115 120 125 Lys Val Ala Ala Ser Cys Gly Ala Ile Gln Tyr Ile Pro ThrGlu Leu 130 135 140 Asp Gln Val Arg Lys Trp Ile Ser Asp Trp Asn Leu ThrThr Glu Lys 145 150 155 160 Lys His Thr Leu Leu Arg Leu Leu Tyr Glu AlaLeu Val Asp Cys Lys 165 170 175 Lys Ser Asp Ala Ala Ser Lys Val Met ValGlu Leu Leu Gly Ser Tyr 180 185 190 Thr Glu Asp Asn Ala Ser Gln Ala ArgVal Asp Ala His Arg Cys Ile 195 200 205 Val Arg Ala Leu Lys Asp Pro AsnAla Phe Leu Phe Asp His Leu Leu 210 215 220 Thr Leu Lys Pro Val Lys PheLeu Glu Gly Glu Leu Ile His Asp Leu 225 230 235 240 Leu Thr Ile Phe ValSer Ala Lys Leu Ala Ser Tyr Val Lys Phe Tyr 245 250 255 Gln Asn Asn LysAsp Phe Ile Asp Ser Leu Gly Leu Leu His Glu Gln 260 265 270 Asn Met AlaLys Met Arg Leu Leu Thr Phe Met Gly Met Ala Val Glu 275 280 285 Asn LysGlu Ile Ser Phe Asp Thr Met Gln Gln Glu Leu Gln Ile Gly 290 295 300 AlaAsp Asp Val Glu Ala Phe Val Ile Asp Ala Val Arg Thr Lys Met 305 310 315320 Val Tyr Cys Lys Ile Asp Gln Thr Gln Arg Lys Val Val Val Ser His 325330 335 Ser Thr His Arg Thr Phe Gly Lys Gln Gln Trp Gln Gln Leu Tyr Asp340 345 350 Thr Leu Asn Ala Trp Lys Gln Asn Leu Asn Lys Val Lys Asn SerLeu 355 360 365 Leu Ser Leu Ser Asp Thr 370 84 13 PRT Homo sapiens 84Met Ser Val Pro Ala Phe Ile Asp Ile Ser Glu Glu Asp 1 5 10 85 15 PRTHomo sapiens 85 Gln Ala Ala Glu Leu Arg Ala Tyr Leu Lys Ser Lys Gly AlaGlu 1 5 10 15 86 17 PRT Homo sapiens 86 Ile Ser Glu Glu Asn Ser Glu GlyGly Leu His Val Asp Leu Ala Gln 1 5 10 15 Ile 87 18 PRT Homo sapiens 87Ile Glu Ala Cys Asp Val Cys Leu Lys Glu Asp Asp Lys Asp Val Glu 1 5 1015 Ser Val 88 16 PRT Homo sapiens 88 Val Ala Arg Pro Ser Ser Leu Phe ArgSer Ala Trp Ser Cys Glu Trp 1 5 10 15 89 12 PRT Homo sapiens 89 Leu ArgLeu Gln Leu Leu Ser Asn Leu Phe His Gly 1 5 10 90 17 PRT Homo sapiens 90Lys Asp Val Glu Ser Val Met Asn Ser Val Val Ser Leu Leu Leu Ile 1 5 1015 Leu 91 26 PRT Homo sapiens 91 Asp Ala Ala Ser Lys Val Met Val Glu LeuLeu Gly Ser Tyr Thr Glu 1 5 10 15 Asp Asn Ala Ser Gln Ala Arg Val AspAla 20 25 92 10 PRT Homo sapiens 92 Val Glu Ala Phe Val Ile Asp Ala ValArg 1 5 10 93 18 PRT Homo sapiens 93 Lys Met Arg Leu Leu Thr Phe Met GlyMet Ala Val Glu Asn Lys Glu 1 5 10 15 Ile Ser 94 196 PRT Homo sapiens 94Met Glu Ala Val Pro Glu Gly Asp Trp Phe Cys Thr Val Cys Leu Ala 1 5 1015 Gln Gln Val Glu Gly Glu Phe Thr Gln Lys Pro Gly Phe Pro Lys Arg 20 2530 Gly Gln Lys Arg Lys Ser Gly Tyr Ser Leu Asn Phe Ser Glu Gly Asp 35 4045 Gly Arg Arg Arg Arg Val Leu Leu Arg Gly Arg Glu Ser Pro Ala Ala 50 5560 Gly Pro Arg Tyr Ser Glu Glu Gly Leu Ser Pro Ser Lys Arg Arg Arg 65 7075 80 Leu Ser Met Arg Asn His His Ser Asp Leu Thr Phe Cys Glu Ile Ile 8590 95 Leu Met Glu Met Glu Ser His Asp Ala Ala Trp Pro Phe Leu Glu Pro100 105 110 Val Asn Pro Arg Leu Val Ser Gly Tyr Arg Arg Ile Ile Lys AsnPro 115 120 125 Met Asp Phe Ser Thr Met Arg Glu Arg Leu Leu Arg Gly GlyTyr Thr 130 135 140 Ser Ser Glu Glu Phe Ala Ala Asp Ala Leu Leu Val PheAsp Asn Cys 145 150 155 160 Gln Thr Phe Asn Glu Asp Asp Ser Glu Val GlyLys Ala Gly His Ile 165 170 175 Met Arg Arg Phe Phe Glu Ser Arg Trp GluGlu Phe Tyr Gln Gly Lys 180 185 190 Gln Ala Asn Leu 195 95 20 PRT Homosapiens 95 Met Glu Ala Val Pro Glu Gly Asp Trp Phe Cys Thr Val Cys LeuAla 1 5 10 15 Gln Gln Val Glu 20 96 21 PRT Homo sapiens 96 Gly Glu PheThr Gln Lys Pro Gly Phe Pro Lys Arg Gly Gln Lys Arg 1 5 10 15 Lys SerGly Tyr Ser 20 97 21 PRT Homo sapiens 97 Leu Asn Phe Ser Glu Gly Asp GlyArg Arg Arg Arg Val Leu Leu Arg 1 5 10 15 Gly Arg Glu Ser Pro 20 98 20PRT Homo sapiens 98 Ala Ala Gly Pro Arg Tyr Ser Glu Glu Gly Leu Ser ProSer Lys Arg 1 5 10 15 Arg Arg Leu Ser 20 99 21 PRT Homo sapiens 99 MetArg Asn His His Ser Asp Leu Thr Phe Cys Glu Ile Ile Leu Met 1 5 10 15Glu Met Glu Ser His 20 100 20 PRT Homo sapiens 100 Asp Ala Ala Trp ProPhe Leu Glu Pro Val Asn Pro Arg Leu Val Ser 1 5 10 15 Gly Tyr Arg Arg 20101 21 PRT Homo sapiens 101 Ile Ile Lys Asn Pro Met Asp Phe Ser Thr MetArg Glu Arg Leu Leu 1 5 10 15 Arg Gly Gly Tyr Thr 20 102 21 PRT Homosapiens 102 Ser Ser Glu Glu Phe Ala Ala Asp Ala Leu Leu Val Phe Asp AsnCys 1 5 10 15 Gln Thr Phe Asn Glu 20 103 17 PRT Homo sapiens 103 Asp AspSer Glu Val Gly Lys Ala Gly His Ile Met Arg Arg Phe Phe 1 5 10 15 Glu104 14 PRT Homo sapiens 104 Ser Arg Trp Glu Glu Phe Tyr Gln Gly Lys GlnAla Asn Leu 1 5 10 105 35 PRT Homo sapiens 105 Met Ser Glu Ile Tyr LeuArg Cys Gln Asp Glu Gln Gln Tyr Ala Arg 1 5 10 15 Trp Met Ala Gly CysArg Leu Ala Ser Lys Gly Arg Thr Met Ala Asp 20 25 30 Ser Ser Tyr 35 10645 PRT Homo sapiens 106 Leu Val Ala Pro Arg Phe Gln Arg Lys Phe Lys AlaLys Gln Leu Thr 1 5 10 15 Pro Arg Ile Leu Glu Ala His Gln Asn Val AlaGln Leu Ser Leu Ala 20 25 30 Glu Ala Gln Leu Arg Phe Ile Gln Ala Trp GlnSer Leu 35 40 45 107 23 PRT Homo sapiens 107 Val Gly Asp Val Val Lys ThrTrp Arg Phe Ser Asn Met Arg Gln Trp 1 5 10 15 Asn Val Asn Trp Asp IleArg 20 108 26 PRT Homo sapiens 108 Glu Glu Ile Asp Cys Thr Glu Glu GluMet Met Val Phe Ala Ala Leu 1 5 10 15 Gln Tyr His Ile Asn Lys Leu SerGln Ser 20 25 109 26 PRT Homo sapiens 109 Glu Glu Ile Asp Cys Thr GluGlu Glu Met Met Val Phe Ala Ala Leu 1 5 10 15 Gln Tyr His Ile Asn LysLeu Ser Gln Ser 20 25 110 26 PRT Homo sapiens 110 Lys Glu Leu Ser PheAla Arg Ile Lys Ala Val Glu Cys Val Glu Ser 1 5 10 15 Thr Gly Arg HisIle Tyr Phe Thr Leu Val 20 25 111 17 PRT Homo sapiens 111 Gly Trp AsnAla Gln Ile Thr Leu Gly Leu Val Lys Phe Lys Asn Gln 1 5 10 15 Gln 112217 PRT Homo sapiens SITE (82) Xaa equals any of the naturally occurringL- amino acids 112 Met Val Thr Thr Ile Val Leu Gly Arg Arg Phe Ile GlySer Ile Val 1 5 10 15 Lys Glu Ala Ser Gln Arg Gly Lys Val Ser Leu PheArg Ser Ile Leu 20 25 30 Leu Phe Leu Thr Arg Phe Thr Val Leu Thr Ala ThrGly Trp Ser Leu 35 40 45 Cys Arg Ser Leu Ile His Leu Phe Arg Thr Tyr SerPhe Leu Asn Leu 50 55 60 Leu Phe Leu Cys Tyr Pro Phe Gly Met Tyr Ile ProPhe Leu Gln Leu 65 70 75 80 Asn Xaa Xaa Leu Arg Lys Thr Ser Leu Phe AsnHis Met Ala Ser Met 85 90 95 Gly Pro Arg Glu Ala Val Ser Gly Leu Ala LysSer Arg Asp Tyr Leu 100 105 110 Leu Thr Leu Arg Glu Thr Trp Lys Gln HisXaa Arg Gln Leu Tyr Gly 115 120 125 Pro Asp Ala Met Pro Thr His Ala CysCys Leu Ser Pro Ser Leu Ile 130 135 140 Arg Ser Glu Val Glu Phe Leu LysMet Asp Phe Asn Trp Arg Met Lys 145 150 155 160 Glu Val Leu Val Ser SerMet Leu Ser Ala Tyr Tyr Val Ala Phe Val 165 170 175 Pro Val Trp Phe ValLys Asn Thr His Tyr Tyr Asp Lys Arg Trp Ser 180 185 190 Cys Xaa Thr LeuPro Ala Gly Val His Gln His Leu Arg Asp Pro His 195 200 205 Ala Ala ProAla Ala Cys Gln Leu Leu 210 215 113 26 PRT Homo sapiens 113 Met Val ThrThr Ile Val Leu Gly Arg Arg Phe Ile Gly Ser Ile Val 1 5 10 15 Lys GluAla Ser Gln Arg Gly Lys Val Ser 20 25 114 23 PRT Homo sapiens 114 LeuPhe Arg Ser Ile Leu Leu Phe Leu Thr Arg Phe Thr Val Leu Thr 1 5 10 15Ala Thr Gly Trp Ser Leu Cys 20 115 30 PRT Homo sapiens 115 Arg Ser LeuIle His Leu Phe Arg Thr Tyr Ser Phe Leu Asn Leu Leu 1 5 10 15 Phe LeuCys Tyr Pro Phe Gly Met Tyr Ile Pro Phe Leu Gln 20 25 30 116 30 PRT Homosapiens SITE (3) Xaa equals any of the naturally occurring L- aminoacids 116 Leu Asn Xaa Xaa Leu Arg Lys Thr Ser Leu Phe Asn His Met AlaSer 1 5 10 15 Met Gly Pro Arg Glu Ala Val Ser Gly Leu Ala Lys Ser Arg 2025 30 117 30 PRT Homo sapiens SITE (14) Xaa equals any of the naturallyoccurring L- amino acids 117 Asp Tyr Leu Leu Thr Leu Arg Glu Thr Trp LysGln His Xaa Arg Gln 1 5 10 15 Leu Tyr Gly Pro Asp Ala Met Pro Thr HisAla Cys Cys Leu 20 25 30 118 31 PRT Homo sapiens 118 Ser Pro Ser Leu IleArg Ser Glu Val Glu Phe Leu Lys Met Asp Phe 1 5 10 15 Asn Trp Arg MetLys Glu Val Leu Val Ser Ser Met Leu Ser Ala 20 25 30 119 27 PRT Homosapiens SITE (24) Xaa equals any of the naturally occurring L- aminoacids 119 Tyr Tyr Val Ala Phe Val Pro Val Trp Phe Val Lys Asn Thr HisTyr 1 5 10 15 Tyr Asp Lys Arg Trp Ser Cys Xaa Thr Leu Pro 20 25 120 20PRT Homo sapiens 120 Ala Gly Val His Gln His Leu Arg Asp Pro His Ala AlaPro Ala Ala 1 5 10 15 Cys Gln Leu Leu 20 121 16 PRT Homo sapiens SITE(7) Xaa equals any of the naturally occurring L- amino acids 121 Leu ValLeu Gly Leu Ser Xaa Leu Asn Asn Ser Tyr Asn Phe Ser Phe 1 5 10 15 122 17PRT Homo sapiens 122 His Val Val Ile Gly Ser Gln Ala Glu Glu Gly Gln TyrSer Leu Asn 1 5 10 15 Phe 123 19 PRT Homo sapiens 123 His Asn Cys AsnAsn Ser Val Pro Gly Lys Glu His Pro Phe Asp Ile 1 5 10 15 Thr Val Met124 17 PRT Homo sapiens 124 Phe Ile Lys Tyr Val Leu Ser Asp Lys Glu LysLys Val Phe Gly Ile 1 5 10 15 Val 125 13 PRT Homo sapiens 125 Ile ProMet Gln Val Leu Ala Asn Val Ala Tyr Ile Ile 1 5 10 126 13 PRT Homosapiens 126 Ile Pro Met Gln Val Leu Ala Asn Val Ala Tyr Ile Ile 1 5 10127 15 PRT Homo sapiens 127 Asp Gly Lys Val Ala Val Asn Leu Ala Lys LeuLys Leu Phe Arg 1 5 10 15 128 13 PRT Homo sapiens 128 Ile Arg Glu LysAsn Pro Asp Gly Phe Leu Ser Ala Ala 1 5 10 129 9 PRT Homo sapiens 129Met Met Phe Gly Gly Tyr Glu Thr Ile 1 5 130 24 PRT Homo sapiens 130 TyrArg Asp Glu Ser Ser Ser Glu Leu Ser Val Asp Ser Glu Val Glu 1 5 10 15Phe Gln Leu Tyr Ser Gln Ile His 20 131 136 PRT Homo sapiens 131 Tyr AlaGln Asp Leu Asp Asp Val Ile Arg Glu Glu Glu His Glu Glu 1 5 10 15 LysAsn Ser Gly Asn Ser Glu Ser Ser Ser Ser Lys Pro Asn Gln Lys 20 25 30 LysLeu Ile Val Leu Ser Asp Ser Glu Val Ile Gln Leu Ser Asp Gly 35 40 45 SerGlu Val Ile Thr Leu Ser Asp Glu Asp Ser Ile Tyr Arg Cys Lys 50 55 60 GlyLys Asn Val Arg Val Gln Ala Gln Glu Asn Ala His Gly Leu Ser 65 70 75 80Ser Ser Leu Gln Ser Asn Glu Leu Val Asp Lys Lys Cys Lys Ser Asp 85 90 95Ile Glu Lys Pro Lys Ser Glu Glu Arg Ser Gly Val Ile Arg Glu Val 100 105110 Met Ile Ile Glu Val Ser Ser Ser Glu Glu Glu Glu Ser Thr Ile Ser 115120 125 Glu Gly Asp Asn Val Glu Ser Trp 130 135 132 37 PRT Homo sapiens132 Met Leu Leu Gly Cys Glu Val Asp Asp Lys Asp Asp Asp Ile Leu Leu 1 510 15 Asn Leu Val Gly Cys Glu Asn Ser Val Thr Glu Gly Glu Asp Gly Ile 2025 30 Asn Trp Ser Ile Ser 35 133 18 PRT Homo sapiens 133 Asp Lys Asp IleGlu Ala Gln Ile Ala Asn Asn Arg Thr Pro Gly Arg 1 5 10 15 Trp Thr 134 31PRT Homo sapiens 134 Gln Arg Tyr Tyr Ser Ala Asn Lys Asn Ile Ile Cys ArgAsn Cys Asp 1 5 10 15 Lys Arg Gly His Leu Ser Lys Asn Cys Pro Leu ProArg Lys Val 20 25 30 135 179 PRT Homo sapiens SITE (120) Xaa equals anyof the naturally occurring L- amino acids 135 Arg Arg Cys Phe Leu CysSer Arg Arg Gly His Leu Leu Tyr Ser Cys 1 5 10 15 Pro Ala Pro Leu CysGlu Tyr Cys Pro Val Pro Lys Met Leu Asp His 20 25 30 Ser Cys Leu Phe ArgHis Ser Trp Asp Lys Gln Cys Asp Arg Cys His 35 40 45 Met Leu Gly His TyrThr Asp Ala Cys Thr Glu Ile Trp Arg Gln Tyr 50 55 60 His Leu Thr Thr LysPro Gly Pro Pro Lys Lys Pro Lys Thr Pro Ser 65 70 75 80 Arg Pro Ser AlaLeu Ala Tyr Cys Tyr His Cys Ala Gln Lys Gly His 85 90 95 Tyr Gly His GluCys Pro Glu Arg Glu Val Tyr Asp Pro Ser Pro Val 100 105 110 Ser Pro PheIle Cys Tyr Tyr Xaa Asp Lys Tyr Glu Ile Gln Glu Arg 115 120 125 Glu LysArg Leu Lys Gln Lys Ile Lys Val Xaa Lys Lys Asn Gly Val 130 135 140 IlePro Glu Pro Ser Lys Leu Pro Tyr Ile Lys Ala Ala Asn Glu Asn 145 150 155160 Pro His His Asp Ile Arg Lys Gly Arg Ala Ser Trp Lys Ser Asn Arg 165170 175 Trp Pro Gln 136 416 PRT Homo sapiens 136 Met Ser Phe Pro Pro HisLeu Asn Arg Pro Pro Met Gly Ile Pro Ala 1 5 10 15 Leu Pro Pro Gly IlePro Pro Pro Gln Phe Pro Gly Phe Pro Pro Pro 20 25 30 Val Pro Pro Gly ThrPro Met Ile Pro Val Pro Met Ser Ile Met Ala 35 40 45 Pro Ala Pro Thr ValLeu Val Pro Thr Val Ser Met Val Gly Lys His 50 55 60 Leu Gly Ala Arg LysAsp His Pro Gly Leu Lys Ala Lys Glu Asn Asp 65 70 75 80 Glu Asn Cys GlyPro Thr Thr Thr Val Phe Val Gly Asn Ile Ser Glu 85 90 95 Lys Ala Ser AspMet Leu Ile Arg Gln Leu Leu Ala Lys Cys Gly Leu 100 105 110 Val Leu SerTrp Lys Arg Val Gln Gly Ala Ser Gly Lys Leu Gln Ala 115 120 125 Phe GlyPhe Cys Glu Tyr Lys Glu Pro Glu Ser Thr Leu Arg Ala Leu 130 135 140 ArgLeu Leu His Asp Leu Gln Ile Gly Glu Lys Lys Leu Leu Val Lys 145 150 155160 Val Asp Ala Lys Thr Lys Ala Gln Leu Asp Glu Trp Lys Ala Lys Lys 165170 175 Lys Ala Ser Asn Gly Asn Ala Arg Pro Glu Thr Val Thr Asn Asp Asp180 185 190 Glu Glu Ala Leu Asp Glu Glu Thr Lys Arg Arg Asp Gln Met IleLys 195 200 205 Gly Ala Ile Glu Val Leu Ile Arg Glu Tyr Ser Ser Glu LeuAsn Ala 210 215 220 Pro Ser Gln Glu Ser Asp Ser His Pro Arg Lys Lys LysLys Glu Lys 225 230 235 240 Lys Glu Asp Ile Phe Arg Arg Phe Pro Val AlaPro Leu Ile Pro Tyr 245 250 255 Pro Leu Ile Thr Lys Glu Asp Ile Asn AlaIle Glu Met Glu Glu Asp 260 265 270 Lys Arg Asp Leu Ile Ser Arg Glu IleSer Lys Phe Arg Asp Thr His 275 280 285 Lys Lys Leu Glu Glu Glu Lys GlyLys Lys Glu Lys Glu Arg Gln Glu 290 295 300 Ile Glu Lys Glu Arg Arg GluArg Glu Arg Glu Arg Glu Arg Glu Arg 305 310 315 320 Glu Arg Arg Glu ArgGlu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu 325 330 335 Lys Glu Lys GluArg Glu Arg Glu Arg Glu Arg Asp Arg Asp Arg Asp 340 345 350 Arg Thr LysGlu Arg Asp Arg Asp Arg Asp Arg Glu Arg Asp Arg Asp 355 360 365 Arg AspArg Glu Arg Ser Ser Asp Arg Asn Lys Asp Arg Ile Arg Ser 370 375 380 ArgGlu Lys Ser Arg Asp Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu 385 390 395400 Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu 405410 415 137 43 PRT Homo sapiens 137 Met Ser Phe Pro Pro His Leu Asn ArgPro Pro Met Gly Ile Pro Ala 1 5 10 15 Leu Pro Pro Gly Ile Pro Pro ProGln Phe Pro Gly Phe Pro Pro Pro 20 25 30 Val Pro Pro Gly Thr Pro Met IlePro Val Pro 35 40 138 35 PRT Homo sapiens 138 Met Ser Ile Met Ala ProAla Pro Thr Val Leu Val Pro Thr Val Ser 1 5 10 15 Met Val Gly Lys HisLeu Gly Ala Arg Lys Asp His Pro Gly Leu Lys 20 25 30 Ala Lys Glu 35 13941 PRT Homo sapiens 139 Asn Asp Glu Asn Cys Gly Pro Thr Thr Thr Val PheVal Gly Asn Ile 1 5 10 15 Ser Glu Lys Ala Ser Asp Met Leu Ile Arg GlnLeu Leu Ala Lys Cys 20 25 30 Gly Leu Val Leu Ser Trp Lys Arg Val 35 40140 40 PRT Homo sapiens 140 Gln Gly Ala Ser Gly Lys Leu Gln Ala Phe GlyPhe Cys Glu Tyr Lys 1 5 10 15 Glu Pro Glu Ser Thr Leu Arg Ala Leu ArgLeu Leu His Asp Leu Gln 20 25 30 Ile Gly Glu Lys Lys Leu Leu Val 35 40141 39 PRT Homo sapiens 141 Lys Val Asp Ala Lys Thr Lys Ala Gln Leu AspGlu Trp Lys Ala Lys 1 5 10 15 Lys Lys Ala Ser Asn Gly Asn Ala Arg ProGlu Thr Val Thr Asn Asp 20 25 30 Asp Glu Glu Ala Leu Asp Glu 35 142 40PRT Homo sapiens 142 Glu Thr Lys Arg Arg Asp Gln Met Ile Lys Gly Ala IleGlu Val Leu 1 5 10 15 Ile Arg Glu Tyr Ser Ser Glu Leu Asn Ala Pro SerGln Glu Ser Asp 20 25 30 Ser His Pro Arg Lys Lys Lys Lys 35 40 143 44PRT Homo sapiens 143 Glu Lys Lys Glu Asp Ile Phe Arg Arg Phe Pro Val AlaPro Leu Ile 1 5 10 15 Pro Tyr Pro Leu Ile Thr Lys Glu Asp Ile Asn AlaIle Glu Met Glu 20 25 30 Glu Asp Lys Arg Asp Leu Ile Ser Arg Glu Ile Ser35 40 144 41 PRT Homo sapiens 144 Lys Phe Arg Asp Thr His Lys Lys LeuGlu Glu Glu Lys Gly Lys Lys 1 5 10 15 Glu Lys Glu Arg Gln Glu Ile GluLys Glu Arg Arg Glu Arg Glu Arg 20 25 30 Glu Arg Glu Arg Glu Arg Glu ArgArg 35 40 145 93 PRT Homo sapiens 145 Glu Arg Glu Arg Glu Arg Glu ArgGlu Arg Glu Arg Glu Lys Glu Lys 1 5 10 15 Glu Arg Glu Arg Glu Arg GluArg Asp Arg Asp Arg Asp Arg Thr Lys 20 25 30 Glu Arg Asp Arg Asp Arg AspArg Glu Arg Asp Arg Asp Arg Asp Arg 35 40 45 Glu Arg Ser Ser Asp Arg AsnLys Asp Arg Ile Arg Ser Arg Glu Lys 50 55 60 Ser Arg Asp Arg Glu Arg GluArg Glu Arg Glu Arg Glu Arg Glu Arg 65 70 75 80 Glu Arg Glu Arg Glu ArgGlu Arg Glu Arg Glu Arg Glu 85 90 146 52 PRT Homo sapiens 146 Arg AspArg Asp Arg Asp Arg Glu Arg Ser Ser Asp Arg Asn Lys Asp 1 5 10 15 ArgIle Arg Ser Arg Glu Lys Ser Arg Asp Arg Glu Arg Glu Arg Glu 20 25 30 ArgGlu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu 35 40 45 ArgGlu Arg Glu 50 147 22 PRT Homo sapiens 147 Lys Pro Gln Met Glu Gly ArgLeu Val Gly Gly Gly Gly Ser Phe Ser 1 5 10 15 Ser Arg Gly Arg His Pro 20148 25 PRT Homo sapiens 148 Leu Leu Val Pro Ser Pro Ser Leu Leu Pro AlaVal Ser Ser Tyr His 1 5 10 15 Leu Pro Leu Gly Arg Gly Leu Ile Arg 20 25149 23 PRT Homo sapiens 149 Glu Gln Gly Ser Ala Val Arg Ser Pro Ala PhePro Val Arg Gln Ala 1 5 10 15 Trp Leu Pro Cys Ser Gly Ser 20 150 151 PRTHomo sapiens SITE (123) Xaa equals any of the naturally occurring L-amino acids 150 Met Gly Leu Asn Pro Pro Gly Leu Thr Ser Ala Leu Lys ProGln Met 1 5 10 15 Glu Gly Arg Leu Val Gly Gly Gly Gly Ser Phe Ser SerArg Gly Arg 20 25 30 His Pro Ala Gly Trp Val Leu Pro Gln Pro Cys Leu LeuLeu Ser Pro 35 40 45 Thr Leu Ser Phe Pro Pro Ala Cys Gly Leu Leu Val ProSer Pro Ser 50 55 60 Leu Leu Pro Ala Val Ser Ser Tyr His Leu Pro Leu GlyArg Gly Leu 65 70 75 80 Ile Arg Pro Ala Phe Lys Ile Lys Val Cys Ser LysLeu Thr Val Trp 85 90 95 Cys Ser Leu Pro Ser Pro Ser Arg Trp Arg Cys CysHis Gly Asn Ala 100 105 110 Val Ala Leu Pro Ala Leu Gly Pro Trp Arg XaaTrp Glu Gln Gly Ser 115 120 125 Ala Val Arg Ser Pro Ala Phe Pro Val ArgGln Ala Trp Leu Pro Cys 130 135 140 Ser Gly Ser Leu Thr Ser Trp 145 150151 64 PRT Homo sapiens 151 Asn Val Thr Lys Ile Thr Leu Glu Ser Phe LeuAla Trp Lys Lys Arg 1 5 10 15 Lys Arg Gln Glu Lys Ile Asp Lys Leu GluGln Asp Met Glu Arg Arg 20 25 30 Lys Ala Asp Phe Lys Ala Gly Lys Ala LeuVal Ile Ser Gly Arg Glu 35 40 45 Val Phe Glu Phe Arg Pro Glu Leu Val AsnAsp Asp Asp Glu Glu Ala 50 55 60 152 22 PRT Homo sapiens 152 Glu Arg ArgLys Ala Asp Phe Lys Ala Gly Lys Ala Leu Val Ile Ser 1 5 10 15 Gly ArgGlu Val Phe Glu 20 153 89 PRT Homo sapiens SITE (81) Xaa equals any ofthe naturally occurring L- amino acids 153 Met Cys Asp Glu Leu Pro GlyGlu Gly Arg Trp Glu Pro Gly Gln Asp 1 5 10 15 Arg Lys Leu Cys Leu SerPhe Pro Leu Gly Thr Pro Ala Arg Pro Ile 20 25 30 Lys Ser Val Cys Pro ThrLeu Leu Ser Leu Val Phe Leu Ser Arg Gly 35 40 45 Met Glu Gln Arg Val ArgGlu Ala Val Ala Val Ser Thr Ser Ala Pro 50 55 60 Ala Pro Ser Ala Ser GluPro Phe Leu Ser Trp Gly Met Gly Leu Ala 65 70 75 80 Xaa Phe Ser Phe ProPhe Leu Tyr Leu 85 154 95 PRT Homo sapiens SITE (71) Xaa equals any ofthe naturally occurring L- amino acids 154 Gly Ala Ser Leu Gly Ser SerSer Ser Cys Pro Ser His Ser Trp Trp 1 5 10 15 Gly Gln Arg Ser Val CysArg Glu Thr Ala Ser Pro Leu Pro Arg Trp 20 25 30 Met Leu Tyr Leu Asp GlyLeu Ala Thr Ser His Phe Leu His His Pro 35 40 45 Glu Pro His Leu Leu ProSer Pro Gly Val Phe Thr Arg Leu Cys Cys 50 55 60 His Leu Cys Pro Gly HisXaa Ser Leu Ser Gly Cys Val Met Asn Ser 65 70 75 80 Gln Glu Arg Glu AspGly Ser Gln Gly Lys Ile Gly Ser Ser Ala 85 90 95 155 125 PRT Homosapiens SITE (30) Xaa equals any of the naturally occurring L- aminoacids 155 Thr Ser Val Leu Ser Ser Ser Ser Val Tyr Cys Met Gln Ala ArgLys 1 5 10 15 Leu Ser Val Ser Gln Arg Tyr Arg Lys Gly Lys Glu Lys XaaAla Arg 20 25 30 Pro Ile Pro Gln Glu Arg Lys Gly Ser Asp Ala Glu Gly AlaGly Ala 35 40 45 Glu Val Glu Thr Ala Thr Ala Ser Leu Thr Leu Cys Ser IlePro Leu 50 55 60 Leu Lys Lys Thr Arg Leu Ser Arg Val Gly Gln Thr Leu PheIle Gly 65 70 75 80 Leu Ala Gly Val Pro Ser Gly Lys Leu Arg Gln Ser PheLeu Ser Cys 85 90 95 Pro Gly Ser His Leu Pro Ser Pro Gly Ser Ser Ser HisIle Pro Arg 100 105 110 Gly Lys Xaa Val Leu Gly Arg Gly Gly Ser Lys AlaGly 115 120 125 156 125 PRT Homo sapiens SITE (13) Xaa equals any of thenaturally occurring L- amino acids 156 Ala Leu Val Lys Gly Thr Gly ArgGlu Lys Arg Arg Xaa Gln Gly Pro 1 5 10 15 Ser Pro Lys Lys Gly Arg AlaLeu Met Gln Arg Glu Gln Glu Leu Arg 20 25 30 Trp Arg Arg Pro Leu Pro LeuSer Pro Ser Val Pro Ser Leu Cys Ser 35 40 45 Arg Lys Pro Gly Leu Ala GluTrp Asp Arg Arg Phe Leu Leu Val Trp 50 55 60 Leu Ala Cys Leu Val Glu SerSer Gly Arg Ala Ser Tyr Leu Ala Leu 65 70 75 80 Ala Pro Ile Phe Pro LeuLeu Gly Val His His Thr Ser Arg Glu Gly 85 90 95 Xaa Val Ser Trp Ala GluVal Ala Ala Lys Pro Gly Lys Asn Ser Arg 100 105 110 Ala Gly Lys Gln MetGly Leu Arg Val Met Gln Lys Met 115 120 125 157 32 PRT Homo sapiens 157Ser Phe Pro Leu Gly Thr Pro Ala Arg Pro Ile Lys Ser Val Cys Pro 1 5 1015 Thr Leu Leu Ser Leu Val Phe Leu Ser Arg Gly Met Glu Gln Arg Val 20 2530 158 31 PRT Homo sapiens 158 Thr Ala Ser Pro Leu Pro Arg Trp Met LeuTyr Leu Asp Gly Leu Ala 1 5 10 15 Thr Ser His Phe Leu His His Pro GluPro His Leu Leu Pro Ser 20 25 30 159 31 PRT Homo sapiens 159 Arg Lys GlySer Asp Ala Glu Gly Ala Gly Ala Glu Val Glu Thr Ala 1 5 10 15 Thr AlaSer Leu Thr Leu Cys Ser Ile Pro Leu Leu Lys Lys Thr 20 25 30 160 25 PRTHomo sapiens 160 Gln Arg Glu Gln Glu Leu Arg Trp Arg Arg Pro Leu Pro LeuSer Pro 1 5 10 15 Ser Val Pro Ser Leu Cys Ser Arg Lys 20 25 161 29 PRTHomo sapiens SITE (13) Xaa equals any of the naturally occurring L-amino acids 161 Pro Leu Leu Gly Val His His Thr Ser Arg Glu Gly Xaa ValSer Trp 1 5 10 15 Ala Glu Val Ala Ala Lys Pro Gly Lys Asn Ser Arg Ala 2025 162 73 PRT Homo sapiens 162 Met Ser Val Leu Lys Gly Glu Arg Gln GlnThr Leu Ala Leu Ala Val 1 5 10 15 Leu Ser Val Ala Lys Glu Asn Ala ArgAsp Val Cys Cys Leu Gln Gly 20 25 30 Trp Gln Asp Thr Ser Cys Arg Asp ThrSer Cys Ala Ala Leu Arg Gly 35 40 45 Gly Leu Gln Thr Leu Phe Pro Ala ProVal His Phe Arg Cys Gly Gly 50 55 60 Pro Ala Glu Leu Lys Gly Arg Gly Ser65 70 163 68 PRT Homo sapiens 163 Ala His Ser Phe Thr Thr Pro Glu GluAla Arg Gly Ala Gly Ser Met 1 5 10 15 Gly Cys Arg Phe Pro Phe Lys HisThr His Ser Pro His Pro Arg Arg 20 25 30 Pro Glu Val Gln Gly Ala Trp AlaGly Cys Thr Ser Ala Gly Glu Lys 35 40 45 Ala Glu Pro Pro Pro Ser Arg GluPro Gly Ser Gln Ala Ser Arg Phe 50 55 60 Pro Leu Pro Pro 65 164 25 PRTHomo sapiens 164 Gly Trp Gln Asp Thr Ser Cys Arg Asp Thr Ser Cys Ala AlaLeu Arg 1 5 10 15 Gly Gly Leu Gln Thr Leu Phe Pro Ala 20 25 165 24 PRTHomo sapiens 165 Gly Cys Arg Phe Pro Phe Lys His Thr His Ser Pro His ProArg Arg 1 5 10 15 Pro Glu Val Gln Gly Ala Trp Ala 20 166 81 PRT Homosapiens 166 Pro His Gln Val Glu Gly Arg Leu Gly Thr Met Glu Thr Trp AspSer 1 5 10 15 Ser His Glu Gly Leu Leu His Cys Arg Ile Pro Leu Lys GlySer Trp 20 25 30 Val Gln Glu Pro Ser Cys Gln Tyr Gln Trp Arg Arg Thr ArgCys Met 35 40 45 Gly Ile Pro Pro Ala Thr Ser Gly Trp Pro Cys Arg Ala ProAla Phe 50 55 60 Leu Cys Ala Arg Ala Glu Phe Pro Ala Ser Pro Gly Gly SerThr Asn 65 70 75 80 Phe 167 81 PRT Homo sapiens 167 Leu Val Thr Pro ProSer Gly Gly Glu Thr Gly Asp His Gly Asn Met 1 5 10 15 Gly Gln Leu ProArg Arg Ala Leu Ala Leu Gln Asn Ser Thr Gln Gly 20 25 30 Ile Leu Gly ProGly Ala Glu Leu Pro Val Ser Val Glu Lys Asp Lys 35 40 45 Val His Gly AspPro Ala Ser Asn Ile Arg Met Ala Met Pro Gly Thr 50 55 60 Arg Phe Pro LeuCys Ser Cys Arg Ile Pro Cys Gln Pro Gly Gly Ile 65 70 75 80 His 168 32PRT Homo sapiens 168 Glu Gly Leu Leu His Cys Arg Ile Pro Leu Lys Gly SerTrp Val Gln 1 5 10 15 Glu Pro Ser Cys Gln Tyr Gln Trp Arg Arg Thr ArgCys Met Gly Ile 20 25 30 169 29 PRT Homo sapiens 169 Gln Asn Ser Thr GlnGly Ile Leu Gly Pro Gly Ala Glu Leu Pro Val 1 5 10 15 Ser Val Glu LysAsp Lys Val His Gly Asp Pro Ala Ser 20 25 170 42 PRT Homo sapiens 170Phe Gly Thr Arg Lys Lys Tyr His Leu Cys Met Ile Pro Asn Leu Asp 1 5 1015 Leu Asn Leu Asp Arg Asp Leu Val Leu Pro Asp Val Ser Tyr Gln Val 20 2530 Glu Ser Ser Glu Glu Asp Gln Ser Gln Thr 35 40 171 115 PRT Homosapiens SITE (88) Xaa equals any of the naturally occurring L- aminoacids 171 Phe Leu Leu Ser Leu Gly Ser Leu Val Met Leu Leu Gln Asp LeuVal 1 5 10 15 His Ser Glu Leu Asp Gly Thr Leu His Tyr Thr Val Ala LeuHis Lys 20 25 30 Asp Gly Ile Glu Met Ser Cys Glu Gln Ser Ile Asp Ser ProAsp Phe 35 40 45 His Leu Leu Asp Trp Lys Cys Thr Val Glu Ile His Lys GluLys Lys 50 55 60 Gln Gln Ser Leu Ser Leu Arg Ile His Ser Leu Arg Leu IleLeu Leu 65 70 75 80 Thr Gly Phe His Leu Ile Thr Xaa Ile Trp Lys His GlnIle Ser Ile 85 90 95 Gln Ile Glu Ile Gln Ile Gly Tyr His Thr Gln Met ValPhe Phe Pro 100 105 110 Arg Ala Glu 115 172 26 PRT Homo sapiens 172 ValHis Ser Glu Leu Asp Gly Thr Leu His Tyr Thr Val Ala Leu His 1 5 10 15Lys Asp Gly Ile Glu Met Ser Cys Glu Gln 20 25 173 28 PRT Homo sapiensSITE (23) Xaa equals any of the naturally occurring L- amino acids 173Gln Ser Leu Ser Leu Arg Ile His Ser Leu Arg Leu Ile Leu Leu Thr 1 5 1015 Gly Phe His Leu Ile Thr Xaa Ile Trp Lys His Gln 20 25 174 340 PRTHomo sapiens 174 Met Ala Ala Ala Cys Gly Pro Gly Ala Ala Gly Thr Ala CysSer Ser 1 5 10 15 Ala Cys Ile Cys Phe Cys Asp Arg Gly Pro Cys Leu GlyTrp Asn Asp 20 25 30 Pro Asp Arg Met Leu Leu Arg Asp Val Lys Ala Leu ThrLeu His Tyr 35 40 45 Asp Arg Tyr Thr Thr Ser Arg Ser Trp Ile Pro Ser HisSer Pro Gln 50 55 60 Leu Lys Cys Val Gly Gly Thr Ala Gly Cys Asp Ser TyrThr Pro Lys 65 70 75 80 Val Ile Gln Cys Gln Asn Lys Gly Trp Asp Gly TyrAsp Val Gln Trp 85 90 95 Glu Cys Lys Thr Asp Leu Asp Ile Ala Tyr Lys PheGly Lys Thr Val 100 105 110 Val Ser Cys Glu Gly Tyr Glu Ser Ser Glu AspGln Tyr Val Leu Arg 115 120 125 Gly Ser Cys Gly Leu Glu Tyr Asn Leu AspTyr Thr Glu Leu Gly Leu 130 135 140 Gln Lys Leu Lys Glu Ser Gly Lys GlnHis Gly Phe Ala Ser Phe Ser 145 150 155 160 Asp Tyr Tyr Tyr Lys Trp SerSer Ala Asp Ser Cys Asn Met Ser Gly 165 170 175 Leu Ile Thr Ile Val ValLeu Leu Gly Ile Ala Phe Val Val Tyr Lys 180 185 190 Leu Phe Leu Ser AspGly Gln Tyr Ser Pro Pro Pro Tyr Ser Glu Tyr 195 200 205 Pro Pro Phe SerHis Arg Tyr Gln Arg Phe Thr Asn Ser Ala Gly Pro 210 215 220 Pro Pro ProGly Phe Lys Ser Glu Phe Thr Gly Pro Gln Asn Thr Gly 225 230 235 240 HisGly Ala Thr Ser Gly Phe Gly Ser Ala Phe Thr Gly Gln Gln Gly 245 250 255Tyr Glu Asn Ser Gly Pro Gly Phe Trp Thr Gly Leu Gly Thr Gly Gly 260 265270 Ile Leu Gly Tyr Leu Phe Gly Ser Asn Arg Ala Ala Thr Pro Phe Ser 275280 285 Asp Ser Trp Tyr Tyr Pro Ser Tyr Pro Pro Ser Tyr Pro Gly Thr Trp290 295 300 Asn Arg Ala Tyr Ser Pro Leu His Gly Gly Ser Gly Ser Tyr SerVal 305 310 315 320 Cys Ser Asn Ser Asp Thr Lys Thr Arg Thr Ala Ser GlyTyr Gly Gly 325 330 335 Thr Arg Arg Arg 340 175 24 PRT Homo sapiens 175Ala Cys Ser Ser Ala Cys Ile Cys Phe Cys Asp Arg Gly Pro Cys Leu 1 5 1015 Gly Trp Asn Asp Pro Asp Arg Met 20 176 26 PRT Homo sapiens 176 ThrAla Gly Cys Asp Ser Tyr Thr Pro Lys Val Ile Gln Cys Gln Asn 1 5 10 15Lys Gly Trp Asp Gly Tyr Asp Val Gln Trp 20 25 177 32 PRT Homo sapiens177 Glu Tyr Asn Leu Asp Tyr Thr Glu Leu Gly Leu Gln Lys Leu Lys Glu 1 510 15 Ser Gly Lys Gln His Gly Phe Ala Ser Phe Ser Asp Tyr Tyr Tyr Lys 2025 30 178 28 PRT Homo sapiens 178 Tyr Lys Leu Phe Leu Ser Asp Gly GlnTyr Ser Pro Pro Pro Tyr Ser 1 5 10 15 Glu Tyr Pro Pro Phe Ser His ArgTyr Gln Arg Phe 20 25 179 26 PRT Homo sapiens 179 Glu Asn Ser Gly ProGly Phe Trp Thr Gly Leu Gly Thr Gly Gly Ile 1 5 10 15 Leu Gly Tyr LeuPhe Gly Ser Asn Arg Ala 20 25 180 25 PRT Homo sapiens 180 Asn Arg AlaTyr Ser Pro Leu His Gly Gly Ser Gly Ser Tyr Ser Val 1 5 10 15 Cys SerAsn Ser Asp Thr Lys Thr Arg 20 25 181 124 PRT Homo sapiens SITE (30) Xaaequals any of the naturally occurring L- amino acids 181 Thr Glu Ser GlnMet Lys Cys Phe Leu Gly Asn Ser His Asp Thr Ala 1 5 10 15 Pro Arg HisThr Cys Ser Gly Gln Gly Leu His Gly Gly Xaa Xaa Xaa 20 25 30 Thr Ala ProLeu Arg Ala Leu Gln Gln His Ser Gln Asp Gly Lys Leu 35 40 45 Cys Thr AsnSer Leu Pro Ala Ala Arg Gly Gly Pro His Lys His Val 50 55 60 Val Val ThrVal Val Tyr Ser Val Lys His Trp Lys Pro Thr Glu Arg 65 70 75 80 Ser SerVal Ser Ile Lys Lys Glu Glu Glu Thr Asp Trp Asp Met Asp 85 90 95 Gln LeuSer Lys Gln Arg Thr Thr Tyr Glu Met Lys Ser Gly Ser Ser 100 105 110 GlyVal Gln Thr Glu Glu Leu Arg His Pro Ser Leu 115 120 182 77 PRT Homosapiens SITE (16) Xaa equals any of the naturally occurring L- aminoacids 182 Asn Ala Ser Trp Glu Ile His Met Thr Gln Arg His Val Ile ProXaa 1 5 10 15 Leu Ala Arg Ala Ser Met Xaa Val Xaa Xaa Xaa Gln Arg ProSer Glu 20 25 30 Leu Cys Ser Ser Ile Arg Arg Met Ala Asn Ser Ala Gln IleVal Phe 35 40 45 Pro Leu Pro Val Gly Ala Pro Thr Asn Thr Leu Ser Ser LeuLeu Tyr 50 55 60 Thr Val Leu Asn Thr Gly Asn Gln Gln Lys Glu Ala Val 6570 75 183 30 PRT Homo sapiens 183 Ala Pro Leu Arg Ala Leu Gln Gln HisSer Gln Asp Gly Lys Leu Cys 1 5 10 15 Thr Asn Ser Leu Pro Ala Ala ArgGly Gly Pro His Lys His 20 25 30 184 27 PRT Homo sapiens 184 Arg Ser SerVal Ser Ile Lys Lys Glu Glu Glu Thr Asp Trp Asp Met 1 5 10 15 Asp GlnLeu Ser Lys Gln Arg Thr Thr Tyr Glu 20 25 185 29 PRT Homo sapiens 185Leu Cys Ser Ser Ile Arg Arg Met Ala Asn Ser Ala Gln Ile Val Phe 1 5 1015 Pro Leu Pro Val Gly Ala Pro Thr Asn Thr Leu Ser Ser 20 25 186 17 PRTHomo sapiens 186 Leu Ser Ile Ile Phe Leu Ala Phe Val Ser Ile Asp Arg CysLeu Gln 1 5 10 15 Leu 187 67 PRT Homo sapiens 187 Gly Ser Cys Phe AlaThr Trp Ala Phe Ile Gln Lys Asn Thr Asn His 1 5 10 15 Arg Cys Val SerIle Tyr Leu Ile Asn Leu Leu Thr Ala Asp Phe Leu 20 25 30 Leu Thr Leu AlaLeu Pro Val Lys Ile Val Val Asp Leu Gly Val Ala 35 40 45 Pro Trp Lys LeuLys Ile Phe His Cys Gln Val Thr Ala Cys Leu Ile 50 55 60 Tyr Ile Asn 65188 31 PRT Homo sapiens 188 Lys Asn Thr Asn His Arg Cys Val Ser Ile TyrLeu Ile Asn Leu Leu 1 5 10 15 Thr Ala Asp Phe Leu Leu Thr Leu Ala LeuPro Val Lys Ile Val 20 25 30 189 17 PRT Homo sapiens 189 Lys His Thr ValGlu Thr Arg Ser Val Ala Phe Arg Lys Gln Leu Asn 1 5 10 15 Arg 190 30 PRTHomo sapiens SITE (18) Xaa equals any of the naturally occurring L-amino acids 190 Pro Gln Val Leu His Leu Arg Trp Leu Pro Lys Val Leu GlyTyr Arg 1 5 10 15 Ser Xaa Pro Leu Arg Leu Ala Asp Pro Ser Thr Phe XaaMet 20 25 30 191 131 PRT Homo sapiens 191 Gln Leu Leu Gly Phe Glu GlyAsn Asp Ser Ala Gly Glu Arg Arg Trp 1 5 10 15 Arg Gly Ala Asn Met GlnIle Pro Leu Leu Gln Val Ala Leu Pro Leu 20 25 30 Ser Thr Glu Glu Gly ThrGly Pro Ser Gly Pro Thr Gln Pro Ser Pro 35 40 45 Gln Gly Glu Val Arg PheLeu Arg Ser Pro Arg Met Gly Gly Gln Val 50 55 60 Pro His Trp Glu Trp ArgSer His Ser Leu Pro Trp Val Leu Thr Ser 65 70 75 80 Thr Leu Ser Gly CysGlu Gly Asp Leu Pro Gly Phe Pro His Gln Val 85 90 95 Gln Leu Pro Ala AlaGlu Ser His Thr Leu Asn Thr Gly Leu Leu Arg 100 105 110 Ser Asp Thr GlyGln Phe Thr Pro Cys Leu Lys Leu Ala Phe Glu Arg 115 120 125 Pro Ser Gly130 192 24 PRT Homo sapiens 192 Asn Asp Ser Ala Gly Glu Arg Arg Trp ArgGly Ala Asn Met Gln Ile 1 5 10 15 Pro Leu Leu Gln Val Ala Leu Pro 20 19329 PRT Homo sapiens 193 Pro Ser Pro Gln Gly Glu Val Arg Phe Leu Arg SerPro Arg Met Gly 1 5 10 15 Gly Gln Val Pro His Trp Glu Trp Arg Ser HisSer Leu 20 25 194 27 PRT Homo sapiens 194 His Gln Val Gln Leu Pro AlaAla Glu Ser His Thr Leu Asn Thr Gly 1 5 10 15 Leu Leu Arg Ser Asp ThrGly Gln Phe Thr Pro 20 25 195 60 PRT Homo sapiens 195 Ala Pro Leu GluThr Met Gln Asn Lys Pro Arg Ala Pro Gln Lys Arg 1 5 10 15 Ala Leu ProPhe Pro Glu Leu Glu Leu Arg Asp Tyr Ala Ser Val Leu 20 25 30 Thr Arg TyrSer Leu Gly Leu Arg Asn Lys Glu Pro Ser Leu Gly His 35 40 45 Arg Trp GlyThr Gln Lys Leu Gly Arg Ser Pro Cys 50 55 60 196 217 PRT Homo sapiensSITE (85) Xaa equals any of the naturally occurring L- amino acids 196Met Gln Asn Lys Pro Arg Ala Pro Gln Lys Arg Ala Leu Pro Phe Pro 1 5 1015 Glu Leu Glu Leu Arg Asp Tyr Ala Ser Val Leu Thr Arg Tyr Ser Leu 20 2530 Gly Leu Arg Asn Lys Glu Pro Ser Leu Gly His Arg Trp Gly Thr Gln 35 4045 Lys Leu Gly Arg Ser Pro Cys Ser Glu Gly Ser Gln Gly His Thr Thr 50 5560 Asp Ala Ala Asp Val Gln Asn His Ser Lys Glu Glu Gln Arg Asp Ala 65 7075 80 Gly Ala Gln Arg Xaa Cys Gly Gln Gly Arg His Thr Trp Ala Tyr Arg 8590 95 Xaa Gly Ala Gln Asp Thr Ser Arg Leu Thr Gly Asp Pro Arg Gly Gly100 105 110 Glu Arg Ser Pro Pro Lys Cys Gln Ser Met Lys Gln Gln Glu GlyAla 115 120 125 Pro Ser Gly His Cys Trp Asp Gln Trp Cys His Gly Ala SerGlu Val 130 135 140 Val Trp Pro Glu Ser Arg Lys Arg Ala Gln Ile Phe XaaSer Pro Cys 145 150 155 160 Arg Gln Ser Pro Arg Ser Ser Ala Leu Gly AlaGly Gln Lys Leu Ala 165 170 175 Val Cys Ser Pro Asp Ile Leu Cys Cys ProThr Asp Thr Leu Leu Ala 180 185 190 Ser His Pro His Ser Leu Leu Thr GlyThr Gln Phe Ser Gly Gln Thr 195 200 205 Gln Ala Leu Ala Pro Ser Trp CysAla 210 215 197 26 PRT Homo sapiens 197 Ala Pro Gln Lys Arg Ala Leu ProPhe Pro Glu Leu Glu Leu Arg Asp 1 5 10 15 Tyr Ala Ser Val Leu Thr ArgTyr Ser Leu 20 25 198 27 PRT Homo sapiens 198 Ala Pro Gln Lys Arg AlaLeu Pro Phe Pro Glu Leu Glu Leu Arg Asp 1 5 10 15 Tyr Ala Ser Val LeuThr Arg Tyr Ser Leu Gly 20 25 199 29 PRT Homo sapiens 199 Leu Gly ArgSer Pro Cys Ser Glu Gly Ser Gln Gly His Thr Thr Asp 1 5 10 15 Ala AlaAsp Val Gln Asn His Ser Lys Glu Glu Gln Arg 20 25 200 25 PRT Homosapiens 200 Thr Asp Thr Leu Leu Ala Ser His Pro His Ser Leu Leu Thr GlyThr 1 5 10 15 Gln Phe Ser Gly Gln Thr Gln Ala Leu 20 25 201 77 PRT Homosapiens SITE (13) Xaa equals any of the naturally occurring L- aminoacids 201 Ile Ala Gln Val Leu Lys Ala Glu Met Cys Leu Val Xaa Arg ProHis 1 5 10 15 Pro Xaa Leu Leu Asp Ser His Arg Gly Trp Ala Gly Glu ThrLeu Arg 20 25 30 Gly Gln Gly Arg Gln Glu Xaa Glu Ser Asp Thr Lys Ala GlyThr Leu 35 40 45 Gln Leu Gln Arg Gln Ala Pro Leu Pro Leu Thr Gln His SerLeu Val 50 55 60 Leu Pro Ile Ser Pro Gly Pro Ser Asn His Thr Gln Ser 6570 75 202 20 PRT Homo sapiens SITE (16) Xaa equals any of the naturallyoccurring L- amino acids 202 Arg Gly Trp Ala Gly Glu Thr Leu Arg Gly GlnGly Arg Gln Glu Xaa 1 5 10 15 Glu Ser Asp Thr 20 203 20 PRT Homo sapiens203 Ala Pro Leu Pro Leu Thr Gln His Ser Leu Val Leu Pro Ile Ser Pro 1 510 15 Gly Pro Ser Asn 20 204 166 PRT Homo sapiens 204 Asn Arg Glu ArgGly Gly Ala Gly Ala Thr Phe Glu Cys Asn Ile Cys 1 5 10 15 Leu Glu ThrAla Arg Glu Ala Val Val Ser Val Cys Gly His Leu Tyr 20 25 30 Cys Trp ProCys Leu His Gln Trp Leu Glu Thr Arg Pro Glu Arg Gln 35 40 45 Glu Cys ProVal Cys Lys Ala Gly Ile Ser Arg Glu Lys Val Val Pro 50 55 60 Leu Tyr GlyArg Gly Ser Gln Lys Pro Gln Asp Pro Arg Leu Lys Thr 65 70 75 80 Pro ProArg Pro Gln Gly Gln Arg Pro Ala Pro Glu Ser Arg Gly Gly 85 90 95 Phe GlnPro Phe Gly Asp Thr Gly Gly Phe His Phe Ser Phe Gly Val 100 105 110 GlyAla Phe Pro Phe Gly Phe Phe Thr Thr Val Phe Asn Ala His Glu 115 120 125Pro Phe Arg Arg Gly Thr Gly Val Asp Leu Gly Gln Gly His Pro Ala 130 135140 Ser Ser Trp Gln Asp Ser Leu Phe Leu Phe Leu Ala Ile Phe Phe Phe 145150 155 160 Phe Trp Leu Leu Ser Ile 165 205 149 PRT Homo sapiens 205 AsnArg Glu Arg Gly Gly Ala Gly Ala Thr Phe Glu Cys Asn Ile Cys 1 5 10 15Leu Glu Thr Ala Arg Glu Ala Val Val Ser Val Cys Gly His Leu Tyr 20 25 30Cys Trp Pro Cys Leu His Gln Trp Leu Glu Thr Arg Pro Glu Arg Gln 35 40 45Glu Cys Pro Val Cys Lys Ala Gly Ile Ser Arg Glu Lys Val Val Pro 50 55 60Leu Tyr Gly Arg Gly Ser Gln Lys Pro Gln Asp Pro Arg Leu Lys Thr 65 70 7580 Pro Pro Arg Pro Gln Gly Gln Arg Pro Ala Pro Glu Ser Arg Gly Gly 85 9095 Phe Gln Pro Phe Gly Asp Thr Gly Gly Phe His Phe Ser Phe Gly Val 100105 110 Gly Ala Phe Pro Phe Gly Phe Phe Thr Thr Val Phe Asn Ala His Glu115 120 125 Pro Phe Arg Arg Gly Thr Gly Val Asp Leu Gly Gln Gly His ProAla 130 135 140 Ser Ser Trp Gln Asp 145 206 41 PRT Homo sapiens 206 AsnArg Glu Arg Gly Gly Ala Gly Ala Thr Phe Glu Cys Asn Ile Cys 1 5 10 15Leu Glu Thr Ala Arg Glu Ala Val Val Ser Val Cys Gly His Leu Tyr 20 25 30Cys Trp Pro Cys Leu His Gln Trp Leu 35 40 207 38 PRT Homo sapiens 207Glu Thr Arg Pro Glu Arg Gln Glu Cys Pro Val Cys Lys Ala Gly Ile 1 5 1015 Ser Arg Glu Lys Val Val Pro Leu Tyr Gly Arg Gly Ser Gln Lys Pro 20 2530 Gln Asp Pro Arg Leu Lys 35 208 34 PRT Homo sapiens 208 Thr Pro ProArg Pro Gln Gly Gln Arg Pro Ala Pro Glu Ser Arg Gly 1 5 10 15 Gly PheGln Pro Phe Gly Asp Thr Gly Gly Phe His Phe Ser Phe Gly 20 25 30 Val Gly209 36 PRT Homo sapiens 209 Ala Phe Pro Phe Gly Phe Phe Thr Thr Val PheAsn Ala His Glu Pro 1 5 10 15 Phe Arg Arg Gly Thr Gly Val Asp Leu GlyGln Gly His Pro Ala Ser 20 25 30 Ser Trp Gln Asp 35 210 15 PRT Homosapiens 210 Gly Leu Ser Thr Gly Pro Asp Met Ala Ser Leu Asp Leu Phe Val1 5 10 15 211 97 PRT Homo sapiens 211 Gly Arg Pro Thr Arg Pro Ser GlnAla Thr Arg His Phe Leu Leu Gly 1 5 10 15 Thr Leu Phe Thr Asn Cys LeuCys Gly Thr Phe Cys Phe Pro Cys Leu 20 25 30 Gly Cys Gln Val Ala Ala AspMet Asn Glu Cys Cys Leu Cys Gly Thr 35 40 45 Ser Val Ala Met Arg Thr LeuTyr Arg Thr Arg Tyr Gly Ile Pro Gly 50 55 60 Ser Ile Cys Asp Asp Tyr MetAla Thr Leu Cys Cys Pro His Cys Thr 65 70 75 80 Leu Cys Gln Ile Lys ArgAsp Ile Asn Arg Arg Arg Ala Met Arg Thr 85 90 95 Phe 212 146 PRT Homosapiens 212 Ile Lys Asn Leu Ile Phe Phe Met Pro Ser Val Val Leu Lys HisIle 1 5 10 15 His His Ile Ser Val Ala Lys Asp Gly Glu Glu Leu Lys LeuLys Arg 20 25 30 Cys Leu Leu Asn Phe Val Ala Ser Val Arg Ala Phe His HisGln Phe 35 40 45 Leu Glu Ser Thr His Gly Ser Pro Ser Val Asp Ile Ser LeuAsp Leu 50 55 60 Ala Lys Ser Thr Met Arg Thr Ala Lys Ser Cys His Ile ValIle Thr 65 70 75 80 Asn Arg Ser Arg Asp Ala Ile Ser Gly Pro Val Glu SerPro His Cys 85 90 95 Asp Ala Cys Ser Thr Gln Thr Ala Phe Ile His Ile SerCys Asn Leu 100 105 110 Thr Pro Lys Ala Arg Glu Thr Lys Cys Ala Thr GluThr Ile Ser Lys 115 120 125 Gln Gly Ser Glu Gln Glu Met Ser Cys Gly LeuGly Arg Thr Arg Gly 130 135 140 Ser Thr 145 213 23 PRT Homo sapiens 213Phe Leu Leu Gly Thr Leu Phe Thr Asn Cys Leu Cys Gly Thr Phe Cys 1 5 1015 Phe Pro Cys Leu Gly Cys Gln 20 214 24 PRT Homo sapiens 214 Ser IleCys Asp Asp Tyr Met Ala Thr Leu Cys Cys Pro His Cys Thr 1 5 10 15 LeuCys Gln Ile Lys Arg Asp Ile 20 215 30 PRT Homo sapiens 215 Ser Val ValLeu Lys His Ile His His Ile Ser Val Ala Lys Asp Gly 1 5 10 15 Glu GluLeu Lys Leu Lys Arg Cys Leu Leu Asn Phe Val Ala 20 25 30 216 26 PRT Homosapiens 216 Asn Phe Val Ala Ser Val Arg Ala Phe His His Gln Phe Leu GluSer 1 5 10 15 Thr His Gly Ser Pro Ser Val Asp Ile Ser 20 25 217 28 PRTHomo sapiens 217 Thr Ala Phe Ile His Ile Ser Cys Asn Leu Thr Pro Lys AlaArg Glu 1 5 10 15 Thr Lys Cys Ala Thr Glu Thr Ile Ser Lys Gln Gly 20 25218 6 PRT Homo sapiens 218 Met Lys Gly Glu Ile Glu 1 5 219 14 PRT Homosapiens 219 Glu Phe Gly Thr Ser Arg Gly Arg Gln His Arg Ala Leu Glu 1 510 220 80 PRT Homo sapiens SITE (72) Xaa equals any of the naturallyoccurring L- amino acids 220 His Gln Thr Pro Gly Val Thr Gly Leu Ser AlaVal Glu Met Asp Gln 1 5 10 15 Ile Thr Pro Ala Leu Trp Glu Ala Leu AlaIle Asp Thr Leu Arg Lys 20 25 30 Leu Arg Ile Gly Thr Arg Arg Pro Arg IleArg Trp Gly Gln Glu Ala 35 40 45 His Val Pro Ala Gly Ala Ala Gln Glu GlyPro Leu His Leu Leu Leu 50 55 60 Gln Arg Pro Ala Pro Trp Gly Xaa Ala ProHis Gly Lys Ala Cys Gly 65 70 75 80 221 87 PRT Homo sapiens SITE (39)Xaa equals any of the naturally occurring L- amino acids 221 Gly Leu GlyGln Gly Gly Gln Gly Leu Asp Gly Gly Arg Lys Leu Met 1 5 10 15 Tyr LeuGln Glu Leu Pro Arg Arg Asp His Tyr Ile Phe Tyr Cys Lys 20 25 30 Asp GlnHis His Gly Gly Xaa Leu His Met Gly Lys Leu Val Gly Arg 35 40 45 Asn SerAsp Thr Asn Arg Glu Ala Leu Glu Glu Phe Lys Lys Leu Val 50 55 60 Gln ArgLys Gly Leu Ser Glu Glu Asp Ile Phe Thr Pro Leu Gln Thr 65 70 75 80 GlySer Cys Val Pro Glu His 85 222 176 PRT Homo sapiens SITE (62) Xaa equalsany of the naturally occurring L- amino acids 222 Ser Gly Pro Ser ArgLeu Arg Thr Ser Leu Ser His Pro Val Ser Asp 1 5 10 15 Val Arg Ala ThrSer Pro Pro Gly Arg Arg Gly Gln Pro Leu Leu Gly 20 25 30 Gly Gly Gln SerTrp Gly Pro Gly Lys Arg Ala Ala Trp Ala Leu Ser 35 40 45 Thr Cys Gly GlyTrp Cys Thr Gly Val Gly Gly Gly Gly Xaa Trp Gly 50 55 60 Trp Glu Trp GlyArg Gly Ser Gln Ala Leu Tyr Leu Pro Gly Ser Ser 65 70 75 80 Val Phe ArgXaa Arg Ile Phe Phe Trp Met His Arg Ser Ser Leu Met 85 90 95 Lys Val AsnVal Ala Ser Asn Phe Pro Pro Pro Arg Ala Val Thr Phe 100 105 110 Thr GlyAsp Thr Phe Trp Ala Ser Cys Leu Arg Lys Val Leu Ser Thr 115 120 125 ThrMet Ala Phe Thr Tyr Gln Val Pro Val Ile Ser Ser Ser Xaa Arg 130 135 140Val Lys Asp Arg Ala Ala Ala Xaa Pro Ser Val Thr Pro Arg Asn Arg 145 150155 160 Val Phe Ile Ser Arg Ala Leu Cys Cys Arg Pro Arg Leu Val Pro Asn165 170 175 223 103 PRT Homo sapiens SITE (74) Xaa equals any of thenaturally occurring L- amino acids 223 Gly Leu Pro Glu Gly Arg Arg AspLeu Val His Leu Asp Cys Gly Gln 1 5 10 15 Ala Cys His Thr Arg Cys LeuMet Ser Gly Pro Pro Ala Pro Gln Glu 20 25 30 Gly Glu Ala Ser Pro Ser LeuGlu Val Gly Arg Ala Gly Ala Leu Ala 35 40 45 Lys Gly Gln Pro Gly His SerLeu Pro Val Glu Ala Gly Ala Leu Gly 50 55 60 Leu Ala Val Gly Glu Gly GlyGly Gly Xaa Gly Gly Gly Ala His Arg 65 70 75 80 Arg Cys Ile Cys Gln AlaPro Pro Ser Ser Ala Xaa Gly Phe Ser Ser 85 90 95 Gly Cys Thr Asp Pro ProSer 100 224 30 PRT Homo sapiens 224 Val Glu Met Asp Gln Ile Thr Pro AlaLeu Trp Glu Ala Leu Ala Ile 1 5 10 15 Asp Thr Leu Arg Lys Leu Arg IleGly Thr Arg Arg Pro Arg 20 25 30 225 23 PRT Homo sapiens 225 Arg Lys LeuMet Tyr Leu Gln Glu Leu Pro Arg Arg Asp His Tyr Ile 1 5 10 15 Phe TyrCys Lys Asp Gln His 20 226 23 PRT Homo sapiens 226 Glu Ala Leu Glu GluPhe Lys Lys Leu Val Gln Arg Lys Gly Leu Ser 1 5 10 15 Glu Glu Asp IlePhe Thr Pro 20 227 27 PRT Homo sapiens 227 Arg Ala Thr Ser Pro Pro GlyArg Arg Gly Gln Pro Leu Leu Gly Gly 1 5 10 15 Gly Gln Ser Trp Gly ProGly Lys Arg Ala Ala 20 25 228 29 PRT Homo sapiens 228 Phe Phe Trp MetHis Arg Ser Ser Leu Met Lys Val Asn Val Ala Ser 1 5 10 15 Asn Phe ProPro Pro Arg Ala Val Thr Phe Thr Gly Asp 20 25 229 28 PRT Homo sapiens229 Cys Leu Met Ser Gly Pro Pro Ala Pro Gln Glu Gly Glu Ala Ser Pro 1 510 15 Ser Leu Glu Val Gly Arg Ala Gly Ala Leu Ala Lys 20 25

What is claimed is:
 1. An isolated antibody or fragment thereof thatspecifically binds to a protein selected from the group consisting of:(a) a protein consisting of amino acid residues 1 to 113 of SEQ IDNO:59; (b) a protein consisting of amino acid residues 30 to 113 of SEQID NO:59; (c) a protein consisting of a portion of SEQ ID NO:59, whereinsaid portion comprises at least 30 contiguous amino acid residues of SEQID NO:59; and (d) a protein consisting of a portion of SEQ ID NO:59,wherein said portion comprises at least 50 contiguous amino acidresidues of SEQ ID NO:59.
 2. The antibody or fragment thereof of claim 1that specifically binds protein (a).
 3. The antibody or fragment thereofof claim 1 that specifically binds protein (b).
 4. The antibody orfragment thereof of claim 1 that specifically binds protein (c).
 5. Theantibody or fragment thereof of claim 1 that specifically binds protein(d).
 6. The antibody or fragment thereof of claim 2 that specificallybinds protein (b).
 7. The antibody or fragment thereof of claim 3,wherein said protein bound by said antibody or fragment thereof isglycosylated.
 8. The antibody or fragment thereof of claim 3 which is ahuman antibody.
 9. The antibody or fragment thereof of claim 3 which isa polyclonal antibody.
 10. The antibody or fragment thereof of claim 3which is selected from the group consisting of: (a) a chimeric antibody;(b) a humanized antibody; (c) a single chain antibody; and (d) a Fabfragment.
 11. The antibody or fragment thereof of claim 3 which islabeled.
 12. The antibody or fragment thereof of claim 11, wherein thelabel is selected from the group consisting of: (a) an enzyme; (b) aradioisotope; and (c) a fluorescent label.
 13. The antibody or fragmentthereof of claim 3, wherein said antibody or fragment thereofspecifically binds to said protein in a Western blot.
 14. The antibodyor fragment thereof of claim 3, wherein said antibody or fragmentthereof specifically binds to said protein in an ELISA.
 15. An isolatedcell that produces the antibody or fragment thereof of claim
 3. 16. Ahybridoma that produces the antibody or fragment thereof of claim
 3. 17.An isolated antibody or fragment thereof obtained from an animal thathas been immunized with a protein selected from the group consisting of:(a) a protein comprising the amino acid sequence of amino acid residues1 to 113 of SEQ ID NO:59; and (b) a protein comprising the amino acidsequence of amino acid residues 30 to 113 of SEQ ID NO:59; (c) a proteincomprising the amino acid sequence of at least 30 contiguous amino acidresidues of SEQ ID NO:59; and (d) a protein comprising the amino acidsequence of at least 50 contiguous amino acid residues of SEQ ID NO:59;wherein said antibody or fragment thereof specifically binds to saidamino acid sequence.
 18. The antibody or fragment thereof of claim 17obtained from an animal immunized with protein (a).
 19. The antibody orfragment thereof of claim 17 obtained from an animal immunized withprotein (b).
 20. The antibody or fragment thereof of claim 17 obtainedfrom an animal immunized with protein (c).
 21. The antibody or fragmentthereof of claim 17 obtained from an animal immunized with protein (d).22. The antibody or fragment thereof of claim 17 which is a monoclonalantibody.
 23. The antibody or fragment thereof of claim 17 which isselected from the group consisting of: (a) a chimeric antibody; (b) apolyclonal antibody; (c) a humanized antibody; (d) a single chainantibody; and (e) a Fab fragment.
 24. An isolated monoclonal antibody orfragment thereof that specifically binds to a protein selected from thegroup consisting of: (a) a protein consisting of amino acid residues 1to 113 of SEQ ID NO:59; (b) a protein consisting of amino acid residues30 to 113 of SEQ ID NO:59; (c) a protein consisting of a portion of SEQID NO:59, wherein said portion comprises at least 30 contiguous aminoacid residues of SEQ ID NO:59; and (d) a protein consisting of a portionof SEQ ID NO:59, wherein said portion comprises at least 50 contiguousamino acid residues of SEQ ID NO:59.
 25. The antibody or fragmentthereof of claim 24 that specifically binds protein (a).
 26. Theantibody or fragment thereof of claim 24 that specifically binds protein(b).
 27. The antibody or fragment thereof of claim 24 that specificallybinds protein (c).
 28. The antibody or fragment thereof of claim 24 thatspecifically binds protein (d).
 29. The antibody or fragment thereof ofclaim 26, wherein said protein bound by said antibody or fragmentthereof is glycosylated.
 30. The antibody or fragment thereof of claim26 which is a human antibody.
 31. The antibody or fragment thereof ofclaim 26 which is selected from the group consisting of: (a) a chimericantibody; (b) a humanized antibody; (c) a single chain antibody; and (d)a Fab fragment.
 32. The antibody or fragment thereof of claim 26 whichis labeled.
 33. The antibody or fragment thereof of claim 32, whereinthe label is selected from the group consisting of: (a) an enzyme; (b) aradioisotope; and (c) a fluorescent label.
 34. The antibody or fragmentthereof of claim 26, wherein said antibody or fragment thereofspecifically binds to said protein in a Western blot.
 35. The antibodyor fragment thereof of claim 26, wherein said antibody or fragmentthereof specifically binds to said protein in an ELISA.
 36. An isolatedcell that produces the antibody or fragment thereof of claim
 26. 37. Ahybridoma that produces the antibody or fragment thereof of claim 26.38. An isolated antibody or fragment thereof that specifically binds toa protein selected from the group consisting of: (a) a proteinconsisting of the full-length polypeptide encoded by the HEMCM42 cDNAcontained in ATCC Deposit Number 209075; (b) a protein consisting of themature form of the polypeptide encoded by the HEMCM42 cDNA contained inATCC Deposit Number 209075; (c) a protein consisting of a portion of thepolypeptide encoded by the HEMCM42 cDNA contained in ATCC Deposit Number209075, wherein said portion comprises at least 30 contiguous amino acidresidues of the polypeptide encoded by the HEMCM42 cDNA contained inATCC Deposit Number 209075; and (d) a protein consisting of a portion ofthe polypeptide encoded by the HEMCM42 cDNA contained in ATCC DepositNumber 209075, wherein said portion comprises at least 50 contiguousamino acid residues of the polypeptide encoded by the HEMCM42 cDNAcontained in ATCC Deposit Number
 209075. 39. The antibody or fragmentthereof of claim 38 that specifically binds protein (a).
 40. Theantibody or fragment thereof of claim 38 that specifically binds protein(b).
 41. The antibody or fragment thereof of claim 38 that specificallybinds protein (c).
 42. The antibody or fragment thereof of claim 38 thatspecifically binds protein (d).
 43. The antibody or fragment thereof ofclaim 39 that specifically binds protein (b).
 44. The antibody orfragment thereof of claim 40, wherein said protein bound by saidantibody or fragment thereof is glycosylated.
 45. The antibody orfragment thereof of claim 40 which is a human antibody.
 46. The antibodyor fragment thereof of claim 40 which is a polyclonal antibody.
 47. Theantibody or fragment thereof of claim 40 which is selected from thegroup consisting of: (a) a chimeric antibody; (b) a humanized antibody;(c) a single chain antibody; and (d) a Fab fragment.
 48. The antibody orfragment thereof of claim 40 which is labeled.
 49. The antibody orfragment thereof of claim 48, wherein the label is selected from thegroup consisting of: (a) an enzyme; (b) a radioisotope; and (c) afluorescent label.
 50. The antibody or fragment thereof of claim 40,wherein said antibody or fragment thereof specifically binds to saidprotein in a Western blot.
 51. The antibody or fragment thereof of claim40, wherein said antibody or fragment thereof specifically binds to saidprotein in an ELISA.
 52. An isolated cell that produces the antibody orfragment thereof of claim
 40. 53. A hybridoma that produces the antibodyor fragment thereof of claim
 40. 54. An isolated antibody or fragmentthereof obtained from an animal that has been immunized with a proteinselected from the group consisting of: (a) a protein comprising theamino acid sequence of the full-length polypeptide encoded by theHEMCM42 cDNA contained in ATCC Deposit Number 209075; (b) a proteincomprising the amino acid sequence of the mature form of the polypeptideencoded by the HEMCM42 cDNA contained in ATCC Deposit Number 209075; (c)a protein comprising the amino acid sequence of at least 30 contiguousamino acid residues of the polypeptide encoded by the HEMCM42 cDNAcontained in ATCC Deposit Number 209075; and (d) a protein comprisingthe amino acid sequence of at least 50 contiguous amino acid residuesthe polypeptide encoded by the HEMCM42 cDNA contained in ATCC DepositNumber 209075; wherein said antibody or fragment thereof specificallybinds to said amino acid sequence.
 55. The antibody or fragment thereofof claim 54 obtained from an animal immunized with protein (a).
 56. Theantibody or fragment thereof of claim 54 obtained from an animalimmunized with protein (b).
 57. The antibody or fragment thereof ofclaim 54 obtained from an animal immunized with protein (c).
 58. Theantibody or fragment thereof of claim 54 obtained from an animalimmunized with protein (d).
 59. The antibody or fragment thereof ofclaim 54 which is a monoclonal antibody.
 60. The antibody or fragmentthereof of claim 54 which is selected from the group consisting of: (a)a chimeric antibody; (b) a polyclonal antibody; (c) a humanizedantibody; (d) a single chain antibody; and (e) a Fab fragment.
 61. Anisolated monoclonal antibody or fragment thereof that specifically bindsto a protein selected from the group consisting of: (a) a proteinconsisting of the full-length polypeptide encoded by the HEMCM42 cDNAcontained in ATCC Deposit Number 209075; (b) a protein consisting of themature form of the polypeptide encoded by the HEMCM42 cDNA contained inATCC Deposit Number 209075; (c) a protein consisting of a portion of thepolypeptide encoded by the HEMCM42 cDNA contained in ATCC Deposit Number209075, wherein said portion comprises at least 30 contiguous amino acidresidues of the polypeptide encoded by the HEMCM42 cDNA contained inATCC Deposit Number 209075; and (d) a protein consisting of a portion ofthe polypeptide encoded by the HEMCM42 cDNA contained in ATCC DepositNumber 209075, wherein said portion comprises at least 50 contiguousamino acid residues of the polypeptide encoded by the HEMCM42 cDNAcontained in ATCC Deposit Number
 209075. 62. The antibody or fragmentthereof of claim 61 that specifically binds protein (a).
 63. Theantibody or fragment thereof of claim 61 that specifically binds protein(b).
 64. The antibody or fragment thereof of claim 61 that specificallybinds protein (c).
 65. The antibody or fragment thereof of claim 61 thatspecifically binds protein (d).
 66. The antibody or fragment thereof ofclaim 62 that specifically binds protein (b).
 67. The antibody orfragment thereof of claim 63, wherein said protein bound by saidantibody or fragment thereof is glycosylated.
 68. The antibody orfragment thereof of claim 63 which is a human antibody.
 69. The antibodyor fragment thereof of claim 63 which is selected from the groupconsisting of: (a) a chimeric antibody; (b) a humanized antibody; (c) asingle chain antibody; and (d) a Fab fragment.
 70. The antibody orfragment thereof of claim 63 which is labeled.
 71. The antibody orfragment thereof of claim 70, wherein the label is selected from thegroup consisting of: (a) an enzyme; (b) a radioisotope; and (c) afluorescent label.
 72. The antibody or fragment thereof of claim 63,wherein said antibody or fragment thereof specifically binds to saidprotein in a Western blot.
 73. The antibody or fragment thereof of claim63, wherein said antibody or fragment thereof specifically binds to saidprotein in an ELISA.
 74. An isolated cell that produces the antibody orfragment thereof of claim
 63. 75. A hybridoma that produces the antibodyor fragment thereof of claim 63.